Cell Culture & Cell Biology - PromoCell

PromoCell GmbH Sickingenstr. 63/65 69126 Heidelberg Germany USA/Canada Phone: 1 – 866 – 251 – 2860 (toll free) Fax: 1 – 866 – 827 – 9219 (toll free) Deutschland Telefon: 0800 – 776 66 23 (gebührenfrei) Fax: 0800 – 100 83 06 (gebührenfrei) France Téléphone: 0800 90 93 32 (ligne verte) Téléfax: 0800 90 27 36 (ligne verte)

2013 | 2014

Cell Culture & Cell Biology

2013 | 2014


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Human Primary Cells & Media Stem and Blood Cells & Media Cell Biology Products

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Overview

1

0

About PromoCell

1

Human Primary Cells

2

Human Stem and Blood Cells

3

Media and Sera

4

Cell Pellets

5

Reagents and Supplements

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Cell Analysis

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Apoptosis

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Fluorescent Labeling

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Cell Transfection

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Antibodies and ELISAs

11

Cytokines and Growth Factors

12

Microbial Detection and Elimination

Contents

Overview

2

Contents

Contents

Contents

0 1

About PromoCell

PromoCell Ethical Standards . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 PromoCell‘s Knowledge Base . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

Human Primary Cells 1.1 Cardiac Myocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.2 Chondrocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.3 Endothelial Cells (Large Vessels) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.4 Endothelial Cells (Microvascular) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.5 Epithelial Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.6 Fibroblasts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.7 Follicle Dermal Papilla Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.8 Hepatocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.9 Keratinocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.10 Melanocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.11 Osteoblasts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.12 Preadipocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.13 Skeletal Muscle Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.14 Smooth Muscle Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

11 12 13 17 23 27 30 31 32 33 34 35 36 37

Overview Human Primary Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42 Human Primary Cells by Tissue Type . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45

2

Human Stem and Blood Cells 2.1 Mesenchymal Stem Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.2 Pericytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.3 CD34+ Progenitor Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.4 CD133+ Progenitor Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.5 Monocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.6 Mononuclear Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

49 51 52 53 54 56

Overview Human Stem and Blood Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58

3 4

Media and Sera 3.1 Media for Primary Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.2 Media for Stem and Blood Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.3 Classical Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.4 Insect Cell Culture Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.5 Sera . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

61 80 85 87 87

Cell Pellets 4.1 Cell Pellets . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91

Contents

6 7 8 9

Reagents and Supplements 5.1 Detachment Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.2 Antibiotics and Antimycotics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.3 Buffers and Salt Solutions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.4 Supplements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

95 97 98 99

Cell Analysis 6.1 Cell Viability and Proliferation Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.2 Live/Dead Cell Staining Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.3 Cytotoxicity Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.4 Senescence Detection Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.5 Cell-Based Reporter Assays . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.6 Cell Staining Reagents and Related Products . . . . . . . . . . . . . . . . . . . . . . . 6.7 Intracellular Indicators and Related Products . . . . . . . . . . . . . . . . . . . . . . . . 6.8 Cell Signaling Assays . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.9 Cell Stress and Metabolism Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.10 Cell Fractionation and Protein Extraction Kits . . . . . . . . . . . . . . . . . . . . . . . 6.11 Protein Size Markers and Protein Gel Stains . . . . . . . . . . . . . . . . . . . . . . . . 6.12 Nucleic Acid Isolation Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.13 DNA Size Markers and DNA Gel Stains . . . . . . . . . . . . . . . . . . . . . . . . . . . .

103 105 106 106 107 109 115 119 120 122 123 124 125

Apoptosis 7.1 DNA Fragmentation Detection Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7.2 Mitochondrial Apoptosis Staining Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7.3 Cytochrome c Apoptosis Detection Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7.4 Glutathione Detection Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7.5 Annexin V Detection Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7.6 Caspase Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7.7 Cathepsin/Calpain Detection Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7.8 Kinase Detection Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7.9 Apoptotic and Necrotic Cell Detection . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7.10 Apoptotic Cell Isolation Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7.11 Apoptosis Inducers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

129 130 130 131 132 134 138 139 140 141 141

Fluorescent Labeling 8.1 8.2 8.3 8.4 8.5

PromoFluor Dyes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . PromoFluor Labeling Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . PromoFluor Labeled Biomolecules . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Phycobiliprotein Dyes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . PromoFluor Antifade Reagent . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

145 148 150 152 153

Cell Transfection 9.1 9.2 9.3 9.4

PromoFectin Transfection Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Magnet Assisted Transfection (MATra) . . . . . . . . . . . . . . . . . . . . . . . . . . . . Transfection Accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Reporter, Expression and Fusion Vectors . . . . . . . . . . . . . . . . . . . . . . . . . . .

157 161 165 166

Contents

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Contents

Contents

Contents

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10.1 Mono- and Polyclonal Antibodies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10.2 Antigens and Cell Lysates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10.3 Labeled Antibodies and Labeling Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10.4 ELISA Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

169 172 173 175

11.1 Cytokines, Growth Factors, Hormones, and soluble Receptors . . . . . . . . . . . 179 11.2 Animal-free Cytokines and Growth Factors . . . . . . . . . . . . . . . . . . . . . . . . . 180 11.3 Cytokines and Growth Factors for Stem Cell Research . . . . . . . . . . . . . . . . 181

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13

Appendix

12.1 Mycoplasma Test Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12.2 Mycoplasma Elimination Solutions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12.3 Bacteria Test Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12.4 Disinfectants . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Alphabetical Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . International Distributors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Terms and Conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Ordering and Technical Support . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Fax Order Form . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

185 186 188 189

191 200 203 205 209

About PromoCell Over 20 years ago PromoCell started its business in the so-called ‚City of Science‘: Heidelberg. Heidelberg is not only home to the oldest university in Germany but is also popular for research related businesses. The city provides the ideal scientific environment for PromoCell and serves as initial point for all our operations.

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6

0 About PromoCell

PromoCell Ethical Standards Ethical and legal standards governing Primary Human Cells

About PromoCell

PromoCell is the original manufacturer of all primary, stem, and blood cells featured in this catalog and is committed to

the highest ethical and legal standards. Consequently, PromoCell strictly complies with the following procedure for the donation and collection of human tissue used for cell isolation:

Extract from “The Convention of Human Rights and Biomedicine”: Article 5 – General rule An intervention in the health field may only be carried out after the person concerned has given free and informed consent to it. This person shall beforehand be given appropriate information as to the purpose and nature of the intervention as well as on its consequences and risks. The person concerned may freely withdraw consent at any time. Article 22 – Disposal of a removed part of the human body When in the course of an intervention any part of a human body is removed, it may be stored and used for a purpose other than that for which it was removed, only if this is done in conformity with appropriate information and consent procedures.

The tissue used by PromoCell for the isolation of human cell cultures is derived from donors who have signed an informed consent form (this being done by the donor himself, an authorized agent, or a legal agent) which outlines in detail the purpose of the donation and the procedure for processing the tissue. We do not accept or use any tissue without prior signing of the consent documents. When obtaining and using human tissue, PromoCell acts in strict compliance with 1. “The Convention for Protection of Human Rights and Dignity of the Human Being with Regard to the Application of Biology and Medicine: Convention of Human Rights and Biomedicine” published on April 4th, 1997 by the Council of Europe (European Treaty Series – no 164). 2. “The Human Tissue Act” published on November 15th, 2004 by the government of the United Kingdom. This act aims to make consent a fundamental principle underpinning the use and storage of human tissue.

Extract from “The Declaration of Helsinki”: Article 22 In any research on human beings, each potential subject must be adequately informed of the aims, methods, sources of funding, any possible conflicts of interest, institutional affiliations of the researcher, the anticipated benefits and potential risks of the study and the discomfort it may entail. The subject should be informed of the right to abstain from participation in the study or to withdraw consent to participate at any time without reprisal. After ensuring that the subject has understood the information, the physician should then obtain the subject‘s freely-given informed consent, preferably in writing. If the consent cannot be obtained in writing, the non-written consent must be formally documented and witnessed.

3. “The Declaration of Helsinki” developed by the World Medical Association as a statement of ethical principles to provide guidance to physicians and other participants in medical research involving human subjects. 4. “The German Federal Data Protection Act” to protect privacy of donors.

Should you have any questions concerning ethical aspects related to the use of Primary Human Cells, do not hesitate to contact us or visit our website: www.promocell.com/ethics

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About PromoCell

0 About PromoCell

At PromoCell we continuously work to improve our products and services.

Dear Customer, Henry Ford once said: ‘Coming together is a beginning. Keeping together is progress. Working together is success.‘ At PromoCell we are proud to work together successfully with scientists worldwide, providing them with high quality products and experienced technical customer support. With over 5,000 PromoCell and PromoKine products we have the tools to meet the needs of your research project! Under our PromoCell brand we offer a broad range of human primary cells, stem cells and blood cells, as well as optimized cell culture media. Scientists worldwide use PromoCell products in basic and applied biomedical research to obtain better results from more accurate physiological models. The reliability of our cells provides the basis for being able to offer you the high quality standards you need. For more in depth analysis of your cells in culture, why not look to our PromoKine brand? PromoKine offers quality kits and reagents for cell biology including those for cell analysis, cell transfection, and fluorescent labeling and also provides numerous antibodies, ELISAs, cytokines and growth factors. In order to complete our portfolio we established the PromoCell Academy in 2004. The PromoCell Academy offers courses with state-of-the-art insight and up to date trends in the Life Sciences in a professional and hands-on setting. You can choose from a growing collection of course topics in German as well as in English. We are looking forward to working with you and providing you with our support! Scientists supporting Scientists – guaranteed.

Your PromoCell Team

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0 About PromoCell

About PromoCell

Discover our Knowledge Base

Need Information?

Information at your fingertips

Sections of the Knowledge Base include:

As scientists, we know that research is not a nine-to-five job. Scientists need information fast, at any time, day or night.

Technical

To assist you with this, PromoCell has created our new online Knowledge Base. At the touch of a button, you can now find a wealth of product-related and technical information to support your research needs.

You can find detailed answers to many questions about our products in the section “Technical Library”. Type in a keyword and the Knowledge Base will show you the associated technical questions and answers within seconds.

Library Notes Reference Literatures Application

In the section “Application Notes” you will find documents which supplement the information in our Product Manuals (e.g. detailed differentiation protocols and staining procedures). If you are interested in viewing publications by our customers, the section “Reference Literature” offers you access to literary citations of scientists that use our PromoCell and PromoKine products.

To access the PromoCell Knowledge Base, please go to: www.promocell.com/knowledge-base

To access the PromoKine Knowledge Base, please go to: www.promokine.info/knowledge-base

We appreciate your feedback We are continually improving our products and services to make them fit your needs. Your feedback, ideas and suggestions therefore are valued and important to us. For your convenience we have created a feedback form online for this purpose: www.promocell.com/feedback

We thank you in advance for providing us with your ideas and suggestions. Of course our customer support team is also happy to receive your feedback via email, fax or telephone. To find your personal area contact, please refer to the contact section on our website or see page 205.

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www.promocell.com/feedback

Human Primary Cells Experiments with human primary cells simulate in vivo conditions much better and yield more significant results than those using immortalized cell lines. This is because the primary cells’ genome and expression patterns have not been altered by immortalization and numerous passages in culture. PromoCell offers a wide variety of high quality human primary cells from tissue obtained from approved medical centers.

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10 Content Chapter 1

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Human Primary Cells 1.1

Cardiac Myocytes . . . . . . . . . . . . . . . . . . . . 11

1.2 Chondrocytes . . . . . . . . . . . . . . . . . . . . . . . 12 1.3

Endothelial Cells (Large Vessels) . . . . . . . . . 13

Umbilical Vein . . . . . . . . . . . . . . . . . . . . . . 13

Umbilical Artery . . . . . . . . . . . . . . . . . . . . . 14

Aortic / Coronary Artery . . . . . . . . . . . . . . . 15

Pulmonary Artery / Saphenous Vein . . . . . . 16

1.4

Endothelial Cells (Microvascular) . . . . . . . . 17

Dermal . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17

Dermal Blood . . . . . . . . . . . . . . . . . . . . . . . 18

Dermal Lymphatic . . . . . . . . . . . . . . . . . . . 19

Bladder / Cardiac . . . . . . . . . . . . . . . . . . . . 20

Pulmonary / Uterine . . . . . . . . . . . . . . . . . 21

Brain . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22 1.5

Epithelial Cells . . . . . . . . . . . . . . . . . . . . . . 23

Nasal / Tracheal . . . . . . . . . . . . . . . . . . . . . 23

Bronchial / Small Airway . . . . . . . . . . . . . . . 24

Mammary / Renal . . . . . . . . . . . . . . . . . . . 25

Renal Cortical / Placental . . . . . . . . . . . . . . 26 1.6 Fibroblasts . . . . . . . . . . . . . . . . . . . . . . . . . 27

Dermal . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27

Pulmonary / Aortic Adventitial / Cardiac . . 28

Uterine / Villous Mesenchymal . . . . . . . . . . 29

1.7

Follicle Dermal Papilla Cells . . . . . . . . . . . . . 30

1.8 Hepatocytes . . . . . . . . . . . . . . . . . . . . . . . . 31 1.9 Keratinocytes . . . . . . . . . . . . . . . . . . . . . . . 32 1.10 Melanocytes . . . . . . . . . . . . . . . . . . . . . . . . 33 1.11 Osteoblasts . . . . . . . . . . . . . . . . . . . . . . . . 34 1.12 Preadipocytes . . . . . . . . . . . . . . . . . . . . . . . 35 1.13 Skeletal Muscle Cells . . . . . . . . . . . . . . . . . 36 1.14 Smooth Muscle Cells . . . . . . . . . . . . . . . . . 37 Aortic . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37

Coronary Artery / Pulmonary Artery . . . . . . 38

Umbilical Artery / Tracheal . . . . . . . . . . . . . 39

Bronchial / Bladder . . . . . . . . . . . . . . . . . . . 40

Prostate / Uterine . . . . . . . . . . . . . . . . . . . . 41

Overview Human Primary Cells . . . . . . . . . . . . . . 42 Human Primary Cells by Tissue Type . . . . . . . . . . . 45

1 Human Primary Cells

11

Human Primary Cells PromoCell offers a wide variety of high quality human primary cells. The tissue used for the isolation of these cells is obtained from approved medical centers following strict ethical standards (see page 6 for details). Shortly after isolation, the cells are cryopreserved using the unique serum-free PromoCell Cryo-SFM (see page 79) and an op-

timized, computer controlled freezing profile. This allows high viability of the cells and defined culture conditions after thawing. All PromoCell primary cells are available cryopreserved or proliferating. Each cell lot is tested for cell type specific markers, cell morphology, population doubling time, and proliferation capacity. In addi-

tion, tests for the absence of HIV 1, HIV 2, HBV, HCV, fungi, mycoplasma, and other bacteria are performed. Moreover, cell pellets prepared from released cell lots and stored in RNAlater® are available. They are useful for DNA, RNA, and protein isolation and their subsequent analysis (for more information see page 91).

Applications Cardiac

research studies Pharmacological studies Physiological

Features Adult

origin α-actinin positive Slow muscle myosin positive Qualified for long-term experiments Sarcomeric

Human Cardiac Myocytes (HCM) are isolated from the ventricles of the adult heart. They are qualified for in vitro research on cardiac diseases and for pharmacological studies. Unlike freshly isolated rod-shaped myocytes, cultured HCM can be used for long-term experiments like investigating the long-term

effects of cytokines, mechanical strain, or cell-cell interactions, as they are prepared according to a special protocol. Initially, the HCM act more like progenitor cells in that they are not yet fully differentiated. They express the markers of early stage differentiation such as GATA-4 and sarcomeric alpha-actin and have a high capacity for proliferation. When they are grown to confluency and cultivated for an extended period of time, the differentiation process begins. Markers of late differentiation (e.g. sarcomeric alpha-actinin, slow muscle myosin) are increased and the cells begin to form myotube-like structures. Recommended media & reagents

Human Primary Cells

1.1 Cardiac Myocytes

Fig. 1: HCM culture in phase contrast. Related products Human Cardiac Microvascular Endothelial Cells (see page 20) Human Cardiac Fibroblasts (see page 28) HCM Pellet (see page 91) PromoFectin Transfection Reagent (see page 157)

Myocyte Growth Medium (see page 62) DetachKit Cryo-SFM

(see page 95) (see page 79)

Count Sarcomeric α-actinin negative

Sarcomeric α-actinin positive Cell Lab Quanta SC TM, Beckman Coulter, Inc.

75

50

Fig. 2: Flow cytometric analysis of PromoCell HCM after being cultured for 60 days under confluent conditions. 96 % of the cells express sarcomeric α-actinin, a marker of late differentiation.

25

0 0

10

102

103

104

Sarcomeric α-actinin

https://www.promocell.com/shop Product Human Cardiac Myocytes (HCM) HCM Pellet

Size

Catalog Number

500,000 cryopreserved cells 500,000 proliferating cells

C-12810 C-12811

1x106 cells in RNAlater®

C-14080

12


1.2 Chondrocytes Applications Osteoarthritis

research research Physiology studies Chondrocyte differentiation experiments Hyaline cartilage investigations Human Primary Cells

Joint

produce and maintain the extracellular matrix of cartilage, i.e. collagen (mostly type II) and proteoglycans (primarily aggrecan).

Recommended media & reagents Growth Medium (see page 62) DetachKit (see page 95) Cryo-SFM (see page 79) Chondrocyte

Related products Pellet (see page 91) PromoFectin Transfection Reagent (see page 157) HCH

Features Available

from hip and knee joints tested by Alcian Blue staining of HCH spheroids Cryopreserved at second passage

Fig. 1: HCH culture in phase contrast.

Human Chondrocytes (HCH) are isolated from normal human articular cartilage from the knee and hip joints (lot specific source information is available on request). PromoCell HCH are routinely tested for their capacity to differentiate in 3D spheroids (see Fig. 2). The articular cartilage covers the joints between bones and contains no blood vessels, lymphatic vessels, or nerve fibers. It is composed of only one specialized cell type, the chondrocytes, which

Fig. 2: Spheroids of differentiated HCH cultured in Chondrocyte Growth Medium and stained with Alcian Blue.

Differentiation

https://www.promocell.com/shop Product Human Chondrocytes (HCH) HCH Pellet

Size

Catalog Number


C-12710 C-12750


C-14070


13

1.3 Endothelial Cells (Large Vessels) Applications Angiogenesis

research research

Atherosclerosis Oncology Drug

uptake studies junction investigation Inflammation studies Cell-cell

Features Von

Willebrand factor (vWF) positive positive Smooth muscle α-actin negative

Tunica media (Smooth muscle cells)

CD31

The endothelium is the thin layer of endothelial cells that line the interior surface of blood and lymphatic vessels - from the aorta to the smallest capillaries. In large vessels (arteries and veins), it is enclosed by two other layers namely the tunica media and the tunica externa (see Fig. 1). PromoCell endothelial cells (large vessels) are available from the umbilical vein and artery, the aorta, the coronary artery, the pulmonary artery, and the saphenous vein. All endothelial cells are tested for positive expression of vWF and CD31, and the absence of smooth muscle α-actin. Available endothelial cell types (large vessels): HUVEC (see below) HUAEC (see page 14) HAoEC (see page 15) HCAEC (see page 15) HPAEC (see page 16) HSaVEC (see page 16)

Human Umbilical Vein Endothelial Cells (HUVEC) HUVEC are isolated from the vein of the umbilical cord and are commonly used for physiological and pharmacological investigations, such as macromolecule transport, blood coagulation, angiogenesis, and fibrinolysis. PromoCell offers HUVEC isolated in standard Endothelial Cell Growth Medium and also in Endothelial Cell

Tunica intima (Endothelial cells)

Fig. 1: Cross section of an artery with the predominant cell types.

Fig. 2: HUVEC culture stained for vWF (Factor VIII antigen). Nuclei counterstained with DAPI.

Growth Medium 2. The cells are supplied either from single donors* or from pooled donors (from up to four different umbilical cords). In addition, HUVEC are available which are pre-screened for VEGF (vascular endothelial growth factor) response. VEGF is an important signaling protein involved in both vasculogenesis and angiogenesis. In vitro, VEGF has been shown to stimulate endothelial cell mitogenesis, cell migration, sprouting, and microvascular permeability.

*Human Umbilical Artery Endothelial Cells (HUAEC) from the same donor are available on request.

Fig. 3: HUVEC culture stained for CD31 antigen. Nuclei counterstained with DAPI.

Recommended media & reagents Cell Growth Medium (see page 63) Endothelial Cell Growth Medium 2 (see page 63) DetachKit (see page 95) Cryo-SFM (see page 79) Endothelial

Human Primary Cells

Tunica externa (Connective tissue)

14


Human Primary Cells

Related products HUVEC single donor Pellet (see page 92) HUVEC pooled Pellet (see page 92) HUVEC Growth Medium 2 single donor Pellet (see page 92) HUVEC Growth Medium 2 pooled Pellet (see page 92) HUVEC pre-screened Pellet (see page 92)

Human

Umbilical Artery Endothelial Cells (see below) PromoFectin-HUVEC Transfection Reagent (see page 158) VEGF (see page 179) CD31 and VEGF Antibodies (see page 169) Fig. 4: HUVEC culture in phase contrast.

https://www.promocell.com/shop Product

Size

Catalog Number


C-12200 C-12250


C-14010


C-12203 C-12253


C-14011

Human Umbilical Vein Endothelial Cells (HUVEC) isolated in Growth Medium 2, single donor


C-12206 C-12207

HUVEC Growth Medium 2 single donor Pellet


C-14008

Human Umbilical Vein Endothelial Cells (HUVEC) isolated in Growth Medium 2, pooled


C-12208 C-12209

HUVEC Growth Medium 2 pooled Pellet


C-14009


C-12205 C-12255


C-14012

Human Umbilical Vein Endothelial Cells (HUVEC) single donor HUVEC single donor Pellet Human Umbilical Vein Endothelial Cells (HUVEC) pooled HUVEC pooled Pellet

Human Umbilical Vein Endothelial Cells (HUVEC) pre-screened HUVEC pre-screened Pellet

Human Umbilical Artery Endothelial Cells (HUAEC) HUAEC are isolated from the arteries of the umbilical cord and are often used for in vitro studies on atherosclerosis but also for endothelial cell-dependent lymphocyte activation and chemoattractant activity experiments*.

Recommended media & reagents Endothelial Cell Growth Medium (see page 63) Endothelial Cell Growth Medium 2 (see page 63) DetachKit (see page 95) Cryo-SFM (see page 79)

Related products Pellet (see page 92) Human Umbilical Vein Endothelial Cells (see page 13) Human Umbilical Artery Smooth Muscle Cells (see page 39) PromoFectin-HUVEC Transfection Reagent (see page 158) CD31 Antibody (see page 169) HUAEC

https://www.promocell.com/shop Product Human Umbilical Artery Endothelial Cells (HUAEC) HUAEC Pellet

Size

Catalog Number


C-12202 C-12252


C-14013

*Human Umbilical Vein Endothelial Cells (HUVEC) from the same donor are available on request.


HAoEC are isolated from the human ascending (thoracic) and descending (abdominal) aorta. They are useful for studying vascular diseases such as thrombosis, atherosclerosis, and hypertension as well as for stent-graft compatibility testing.

Recommended media & reagents Cell Growth Medium MV (see page 64) Endothelial Cell Growth Medium MV2 (see page 64) DetachKit (see page 95) Cryo-SFM (see page 79) Endothelial

Related products HAoEC Pellet (see page 91) Human Aortic Smooth Muscle Cells (see page 37) Human Aortic Adventitial Fibroblasts (see page 28) PromoFectin-HUVEC Transfection Reagent (see page 158) CD31 Antibody (see page 169) https://www.promocell.com/shop

Product Human Aortic Endothelial Cells (HAoEC) HAoEC Pellet

Human Coronary Artery Endothelial Cells (HCAEC) HCAEC are isolated from the coronary arteries (the right and the left coronary artery, including the anterior descending and the circumflex branches) from a

Size

Catalog Number


C-12271 C-12272


C-14023

single donor. They are crucially involved in the regulation of coronary blood flow and cardiac functions and are consequently useful for in vitro studies of diseases such as thrombosis, atherosclerosis, and hypertension. HCAEC are also suitable for studying functional vaso-

dilators, such as prostacyclin, bradykinin, or nitric oxide. Recommended media & reagents Cell Growth Medium MV (see page 64) Endothelial Cell Growth Medium MV2 (see page 64) DetachKit (see page 95) Cryo-SFM (see page 79) Endothelial

Related products Pellet (see page 91) Human Coronary Artery Smooth Muscle Cells (see page 38) PromoFectin-HUVEC Transfection Reagent (see page 158) CD31 Antibody (see page 169) HCAEC

Left main coronary artery

Circumflex coronary artery

Right coronary artery Left anterior descending coronary artery Fig. 5: Location of the coronary arteries.

https://www.promocell.com/shop Product Human Coronary Artery Endothelial Cells (HCAEC) HCAEC Pellet

Size

Catalog Number


C-12221 C-12222


C-14022

Human Primary Cells

Human Aortic Endothelial Cells (HAoEC)

15

16


Human Primary Cells

Human Pulmonary Artery Endothelial Cells (HPAEC) HPAEC are isolated from human pulmonary arteries (the main pulmonary artery, left and right branches). Since the pulmonary arteries carry blood from the heart to the lungs, they are the only arteries (other than umbilical arteries) that carry deoxygenated blood.

Recommended media & reagents Cell Growth Medium (see page 63) Endothelial Cell Growth Medium 2 (see page 63) DetachKit (see page 95) Cryo-SFM (see page 79)

Endothelial

Related products Pellet (see page 92) Human Pulmonary Artery Smooth Muscle Cells (see page 38) PromoFectin-HUVEC Transfection Reagent (see page 158) CD31 Antibody (see page 169) HPAEC

https://www.promocell.com/shop Product Human Pulmonary Artery Endothelial Cells (HPAEC) HPAEC Pellet

Human Saphenous Vein Endothelial Cells (HSaVEC) HSaVEC are isolated from human saphenous veins from single donors. The saphenous vein is often used for autotransplantation in coronary artery bypass operations, when arterial grafts are not available or many grafts are required.

Size

Catalog Number


C-12241 C-12242


C-14024

Recommended media & reagents Cell Growth Medium (see page 63) Endothelial Cell Growth Medium 2 (see page 63) DetachKit (see page 95) Cryo-SFM (see page 79)

Endothelial

Related products Pellet (see page 92) PromoFectin-HUVEC Transfection Reagent (see page 158) CD31 Antibody (see page 169) HSaVEC

https://www.promocell.com/shop Product Human Saphenous Vein Endothelial Cells (HSaVEC) HSaVEC Pellet

Size

Catalog Number


C-12231 C-12232


C-14025


17

1.4 Endothelial Cells (Microvascular) Applications Angiogenesis

research

Oncology Drug

uptake studies junction investigation Inflammation studies

at the interface between the circulating fluid in the lumen and the surrounding tissue, microvascular endothelial cells are highly active and closely involved in numerous physiological processes.

Features Von

Willebrand factor (vWF) positive positive Smooth muscle α-actin negative Cryopreserved at second passage CD31

The walls of capillaries are composed of a single layer of microvascular endothelial cells. This layer is so thin that molecules such as oxygen, water, and lipids can pass through it by diffusion and enter the surrounding tissues. Waste products such as carbon dioxide and urea, on the other hand, diffuse back into the blood to be carried away. Because of their location

Human Dermal Microvascular Endothelial Cells (HDMEC) HDMEC are isolated from the dermis of juvenile foreskin and adult skin (different locations). Since the dermis contains blood and lymphatic capillaries, HDMEC comprise Blood and Lymphatic Microvascular Endothelial Cells. Both have a common origin and can be identified by several markers. In addition, HDMEC are available which are pre-screened for VEGF (vascular endothelial growth factor) response. VEGF is an important signaling protein involved in both vasculogenesis and angiogenesis. In vitro, VEGF has been shown to stimu-

Microvascular endothelial cells differ in morphologies and properties according to the tissues supplied by the capillaries. Therefore, PromoCell microvascular endothelial cells are available from dermal, lung, cardiac, uterine, bladder, and brain tissues. Available endothelial cell types (microvascular): HDMEC (see below) HDBEC (see page 18) HDLEC (see page 19) HBdMEC (see page 20) HCMEC (see page 20) HPMEC (see page 21) HUtMEC (see page 21) HBMEC (see page 22)

late endothelial cell mitogenesis, cell migration, sprouting, and microvascular permeability.

Fig. 1: HDMEC culture stained for vWF (Factor VIII antigen). Nuclei counterstained with DAPI.

Fig. 2: HDMEC culture stained for CD31 antigen. Nuclei counterstained with DAPI.

Endothelial

Pericytes (see page 51) Transfection Reagent (see page 158) VEGF (see page 179) CD31 Antibody (see page 169)

Related products Pellet (see page 91) HDMEC adult Pellet (see page 91) HDMEC pre-screened Pellet (see page 91)

Fig. 3: HDMEC culture in phase contrast.

Recommended media & reagents Cell Growth Medium MV (see page 64) Endothelial Cell Growth Medium MV2 (see page 64) DetachKit (see page 95) Cryo-SFM (see page 79)

Human

PromoFectin-HUVEC

HDMEC

https://www.promocell.com/shop Product Human Dermal Microvascular Endothelial Cells (HDMEC) juvenile foreskin HDMEC Pellet Human Dermal Microvascular Endothelial Cells (HDMEC) adult donor HDMEC adult Pellet Human Dermal Microvascular Endothelial Cells (HDMEC) pre-screened HDMEC pre-screened Pellet

Size

Catalog Number


C-12210 C-12260


C-14015


C-12212 C-12262


C-14016


C-12215 C-12265


C-14017

Human Primary Cells

Cell-cell

18


HDBEC are isolated from the dermis of juvenile foreskin and adult skin (different locations) from a single donor*. The cells are routinely analyzed by flow cytometric analysis and immunofluorescent staining: they are > 95% CD31 positive (see Fig. 4) and stain positive for von Willebrand factor. In addition, they are > 90% negative for podoplanin (see Fig. 5) and stain negative for smooth muscle specific α-actin.

Blood Endothelial Cells have a key function in physiological processes like vessel tonus, capillary permeability, blood coagulation, fibrolysis, and angiogenesis. They are also useful for disease studies of atherosclerosis, tumorigenesis, and thrombosis. Recommended media & reagents Cell Growth Medium MV (see page 64) DetachKit (see page 95) Cryo-SFM (see page 79)

Related products Pellet (see page 91) HDBEC adult Pellet (see page 91) Human Dermal Lymphatic Endothelial Cells (see page 19) PromoFectin-HUVEC Transfection Reagent (see page 158) Podoplanin, CD31, and Prox-1 Antibodies (see page 169) HDBEC

Endothelial

Count 75

CD31 negative

Cell Lab Quanta SC TM, Beckman Coulter, Inc.

CD31 positive

50

25

Fig. 4: Flow cytometric analysis of PromoCell HDBEC. 98% of the cells are CD31 positive .

0 0

10

102

103

104

CD31

Count 75

Podoplanin negative

Podoplanin positive Cell Lab Quanta SC TM, Beckman Coulter, Inc.

Human Primary Cells

Human Dermal Blood Endothelial Cells (HDBEC)

50

25

Fig. 5: Flow cytometric analysis of PromoCell HDBEC. 91% of the cells are podoplanin negative.

0 0

10

102

103

104

Podoplanin

https://www.promocell.com/shop Product Human Dermal Blood Endothelial Cells (HDBEC) juvenile foreskin HDBEC Pellet Human Dermal Blood Endothelial Cells (HDBEC) adult donor HDBEC adult Pellet

Size

Catalog Number


C-12211 C-12214


C-14018


C-12225 C-12226


C-14019

*Human Dermal Lymphatic Microvascular Endothelial Cells (HDLEC) from the same donor are available on request.


HDLEC are isolated from the dermis of juvenile foreskin and adult skin (different locations) from a single donor*. The cells are routinely analyzed by flow cytometric analysis and immunofluorescent staining: > 95% of the cells are CD31 positive and podoplanin positive (see Fig. 6 and 7). In addition, the cells stain positive for Prox-1. Lymphatic vessels transport excess fluids from tissues to the circulatory system and are a major component of the immune

system. The transported fluid is nearly cell free. The vessels are highly permeable, because they lack a continuous basal membrane. Lymphatic endothelial cells are involved in pathological alterations of the lymphatic system. During tumor lymphangiogenesis, the lymphatic endothelial cells build new vessels that infiltrate tumors, attract tumor cells, and induce tumor cell metastasis. Recommended media & reagents Cell Growth Medium MV2 (see page 64) Endothelial

DetachKit Cryo-SFM


Related products Pellet (see page 91) HDLEC adult Pellet (see page 91) Human Dermal Blood Endothelial Cells (see page 18) PromoFectin-HUVEC Transfection Reagent (see page 158) VEGF (see page 179) Podoplanin, CD31, and Prox-1 Antibodies (see page 169) HDLEC

Count 75

CD31 positive Cell Lab Quanta SC TM, Beckman Coulter, Inc.

CD31 negative

50

25

Fig. 6: Flow cytometric analysis of PromoCell HDLEC. 99% of the cells are CD31 positive.

0 0

10

102

103

104

CD31

Count 75

Podoplanin positive Cell Lab Quanta SC T M, Beckman Coulter, Inc.

Podoplanin negative

50

25

Fig. 7: Flow cytometric analysis of PromoCell HDLEC. 96% of the cells are podoplanin positive.

0 0

10

102

103

104

Podoplanin

https://www.promocell.com/shop Product Human Dermal Lymphatic Endothelial Cells (HDLEC) juvenile foreskin HDLEC Pellet Human Dermal Lymphatic Endothelial Cells (HDLEC) adult donor HDLEC adult Pellet

Size

Catalog Number


C-12216 C-12218


C-14020


C-12217 C-12219


C-14021

*Human Dermal Blood Endothelial Cells (HDBEC) from the same donor are available on request.

Human Primary Cells

Human Dermal Lymphatic Endothelial Cells (HDLEC)

19

20


Human Primary Cells

Human Bladder Microvascular Endothelial Cells (HBdMEC) HBdMEC are isolated from the human bladder microvasculature. The cells are routinely analyzed by immunofluorescent staining: they stain positive for CD31 and von Willebrand factor and negative for smooth muscle α-actin. HBdMEC are useful in studying the role of microvascular endothelial cells in the control of inflammation, capillary exchange, tissue growth, and angiogenesis.


Related products Pellet (see page 91) PromoFectin-HUVEC Transfection Reagent (see page 158) CD31 Antibody (see page 169) HBdMEC

https://www.promocell.com/shop Product Human Bladder Microvascular Endothelial Cells (HBdMEC) HBdMEC Pellet

Human Cardiac Microvascular Endothelial Cells (HCMEC) HCMEC are isolated from heart ventricles from single donors. The cells are routinely analyzed by immunofluorescent staining: they stain positive for CD31 and von Willebrand factor and negative for smooth muscle α-actin. HCMEC intensively interact with cardiomyocytes and therefore have a characteristic phenotype different from other microvascular endothelial cells. HCMEC

Size

Catalog Number


C-12291 C-12292


C-14026

play an important role in the physiological regulation of coronary blood flow and capillary exchange. Medical conditions such as hypertrophy, ischemia, hypertension, and diabetes mellitus are associated with endothelial dysfunction. Recommended media & reagents Cell Growth Medium MV (see page 64) Endothelial Cell Growth Medium MV2 (see page 64) DetachKit (see page 95) Cryo-SFM (see page 79) Endothelial

Related products HCMEC Pellet (see page 91) Human Cardiac Myocytes (see page 11) Human Cardiac Fibroblasts (see page 28) PromoFectin-HUVEC Transfection Reagent (see page 158) CD31 Antibody (see page 169)

https://www.promocell.com/shop Product Human Cardiac Microvascular Endothelial Cells (HCMEC) HCMEC Pellet

Size

Catalog Number


C-12285 C-12286


C-14029


HPMEC are isolated from the lung from a single donor. The cells are routinely analyzed by immunofluorescent staining: they stain positive for CD31 and von Willebrand factor and negative for smooth muscle α-actin. The lung has a vast endothelial surface area, which is essential for the exchange of gases. HPMEC are most appropriate for studying human lung diseases.


Related products Pellet (see page 92) Human Pulmonary Fibroblasts (see page 28) Human Small Airway Epithelial Cells (see page 24) PromoFectin-HUVEC Transfection Reagent (see page 158) CD31 Antibody (see page 169) HPMEC

https://www.promocell.com/shop Product Human Pulmonary Microvascular Endothelial Cells (HPMEC) HPMEC Pellet

Human Uterine Microvascular Endothelial Cells (HUtMEC) HUtMEC are isolated from the myometrium (middle layer of the uterine wall) from a single donor, who has not been pre-treated with hormones. The cells are routinely analyzed by immunofluorescent staining: they stain positive for CD31 and von Willebrand factor and negative for smooth muscle α-actin. During pregnancy HUtMEC propagate intensely and change their phenotype and expression pattern in response to sex steroids. This includes up-regulation

Size

Catalog Number


C-12281 C-12282


C-14027

of HUtMEC nitric oxide synthase activity, modulation of adhesion molecule expression, and an increase in endothelial cell survival and angiogenic activity. HUtMEC are involved in medical conditions such as uterine fibroma or adenomyosis. Recommended media & reagents Cell Growth Medium MV (see page 64) Endothelial Cell Growth Medium MV2 (see page 64) DetachKit (see page 95) Cryo-SFM (see page 79) Endothelial

Related products Pellet (see page 92) Human Uterine Fibroblasts (see page 29) Human Uterine Smooth Muscle Cells (see page 41) PromoFectin-HUVEC Transfection Reagent (see page 158) CD31 Antibody (see page 169) HUtMEC

https://www.promocell.com/shop Product Human Uterine Microvascular Endothelial Cells (HUtMEC) HUtMEC Pellet

Size

Catalog Number


C-12295 C-12296


C-14028

Human Primary Cells

Human Pulmonary Microvascular Endothelial Cells (HPMEC)

21

22


Human Primary Cells

Human Brain Microvascular Endothelial Cells (HBMEC) Human Brain Microvascular Endothelial Cells (HBMEC) are isolated from cortical tissue (see Fig. 8) from a single donor. They are routinely analyzed by flow cytometry and immunofluorescent staining. The cells are positive for CD31 and von Willebrand factor and negative for smooth muscle α-actin and CD90.

Brain microvascular endothelial cells are highly specialized cells responsible for the formation and functional properties of the Blood-Brain Barrier (BBB). The BBB is characterized by cellular tight junctions and active transport systems which allow the selective transport of substances into the brain, and prevent the entrance of unwanted substances. Therefore, PromoCell HBMEC are well suited for studying brain endothelial cell

functions like active transport systems and drug uptake. Furthermore, neuroinflammation studies as well as tight junction research can be performed with HBMEC. Recommended media & reagents Cell Growth Medium MV2 (see page 64) DetachKit (see page 95) Cryo-SFM (see page 79)

Endothelial

Cerebral cortex Thalamus Corpus callosum

Hypothalamus Pituitary gland Cerebellum Spinal cord

Fig. 8: Schematic structure of the brain.

Fig. 9: HBMEC in phase contrast.

https://www.promocell.com/shop Product Human Brain Microvascular Endothelial Cells (HBMEC) HBMEC Pellet

Size

Catalog Number


Limited Availability




C-12287 C-12288 C-14032


23

1.5 Epithelial Cells

Drug uptake studies Toxicological research Wound healing research Oncology Cell differentiation studies Respiratory disease research

Typically, epithelial tissue is made of tightly packed cells with little intercellular space and is arranged in flat sheets forming effective barriers between the underlying tissues and the external environment.

Cytokeratin positive Available from different tissues Cryopreserved at second passage

PromoCell offers epithelial cells from different locations: Airways: Nasal mucosa, trachea, bronchi, and distal respiratory tract Breast: Mammary gland Kidney: Whole kidney and renal cortex Placenta: Amniotic membrane

The epithelium is the interface between the body and the external environment. It covers all outside surfaces and inside lumina e.g. the inner lining of the respiratory tract. In addition, epithelial cells also make up exocrine and endocrine glands. The functions of epithelial cells are diverse and include absorption, secretion, protection, transcellular transport, and sensation.

Available Epithelial Cells: HNEpC (see below) HTEpC (see below) HBEpC (see page 24) HSAEpC (see page 24) HMEpC (see page 25) HREpC (see page 25) HRCEpC (see page 26) HPlEpC (see page 26)

Human Nasal Epithelial Cells (HNEpC)

particles that are subsequently transported to the pharynx by cilia on the epithelial cells. HNEpC are useful for in vitro studies of these processes.

Features

HNEpC are isolated from normal human nasal mucosa and stain positive for cytokeratin. The epithelium in the nasal cavity cleans, humidifies, and warms inhaled air. In addition, it produces mucus, which binds

Fig. 1: HTEpC culture in phase contrast. Note the cuboidal shape typical for airway epithelial cells.

Fig. 2: HTEpC culture stained for cytokeratin. Nuclei counterstained with DAPI.

DetachKit Cryo-SFM


Related products Pellet (see page 92) PromoFectin Transfection Reagent (see page 157) HNEpC

Recommended media & reagents Epithelial Cell Growth Medium (see page 65)

Airway

https://www.promocell.com/shop Product Human Nasal Epithelial Cells (HNEpC) HNEpC Pellet

Human Tracheal Epithelial Cells (HTEpC) HTEpC are isolated from the surface epithelium of human trachea and stain positive for cytokeratin. The respiratory epithelia are responsible for the lubrication of the lungs, the maintenance of humidity, and the cleaning of the respiratory tract. They

Size

Catalog Number


C-12620 C-12621


C-14062

are an important target for drugs, toxins, and carcinogens. In addition, they are involved in many diseases such as cystic fibrosis. Consequently, HTEpC are useful for investigating the function and pathology of the respiratory system. Recommended media & reagents Epithelial Cell Growth Medium (see page 65)

Airway

DetachKit Cryo-SFM


Related products Pellet (see page 92) Human Tracheal Smooth Muscle Cells (see page 39) PromoFectin Transfection Reagent (see page 157) HTEpC

Human Primary Cells

Applications

24


https://www.promocell.com/shop Product Human Tracheal Epithelial Cells (HTEpC)

Human Primary Cells

HTEpC Pellet

Human Bronchial Epithelial Cells (HBEpC) HBEpC are isolated from the surface epithelium of human bronchi and stain positive for cytokeratin. The respiratory epithelia are responsible for the lubrication of the lungs, the maintenance of humidity, and the cleaning of the respiratory tract. They

Size

Catalog Number


C-12644 C-12645


C-14064

are an important target for drugs, toxins, and carcinogens. In addition, they are involved in many diseases such as cystic fibrosis. Consequently, HBEpC are useful for investigating the function and pathology of the respiratory system. Recommended media & reagents Epithelial Cell Growth Medium (see page 65)

Airway

DetachKit Cryo-SFM


Related products Pellet (see page 91) Human Bronchial Smooth Muscle Cells (see page 40) PromoFectin Transfection Reagent (see page 157) HBEpC

https://www.promocell.com/shop Product Human Bronchial Epithelial Cells (HBEpC) HBEpC Pellet

Human Small Airway Epithelial Cells (HSAEpC) HSAEpC are isolated from the distal portion of the human respiratory tract in the 1 mm bronchiole area. They stain positive for cytokeratin. The distal respiratory tract mainly consists of pulmonary alveoli, which are spherical outcroppings of the respiratory bronchioles and are the primary sites of gas exchange with the blood. HSAEpC are qualified for functional

Size

Catalog Number


C-12640 C-12641


C-14063

studies to investigate disorders such as asthma or pulmonary inflammation.

Recommended media & reagents Airway Epithelial Cell Growth Medium (see page 66) DetachKit (see page 95) Cryo-SFM (see page 79) Small

Related products Pellet (see page 92) Human Pulmonary Microvascular Endothelial Cells (see page 21) Human Pulmonary Fibroblasts (see page 28) PromoFectin Transfection Reagent (see page 157) HSAEpC

Fig. 3: HSAEpC culture in phase contrast.

https://www.promocell.com/shop Product Human Small Airway Epithelial Cells (HSAEpC) HSAEpC Pellet

Size

Catalog Number


C-12642 C-12643


C-14065


HMEpC are isolated from the human adult mammary gland and stain positive for cytokeratin. Hormones extensively control the development of the mammary glands. Throughout puberty, the glands begin to develop in females in response to ovarian hormones. Further on during pregnancy, rising levels of estrogen and progesterone cause additional growth and differentiation. Finally, due to the presence of the hormone prolactin in late pregnancy,

milk secretion (lactation) begins. Since mammary gland cells can easily be induced to grow and multiply by hormones, HMEpC are very well suited for developmental investigations. In addition, they are useful for breast cancer in vitro research since breast cancer usually originates in the lobules or ducts of the mammary glands. Recommended media & reagents Epithelial Cell Growth Medium (see page 67) DetachKit (see page 95) Cryo-SFM (see page 79)

Related products Pellet (see page 91) PromoFectin Transfection Reagent (see page 157) HMEpC

Human Primary Cells

Human Mammary Epithelial Cells (HMEpC)

25

Mammary

Fig. 4: HMEpC culture in phase contrast.

https://www.promocell.com/shop Product Human Mammary Epithelial Cells (HMEpC) HMEpC Pellet

Human Renal Epithelial Cells (HREpC) HREpC are isolated from the adult kidney and stain positive for cytokeratin. Kidneys have a very complex structure and contain diverse epithelia such as those in the convoluted tubules, the cortical collecting ducts, the calyxes,

Renal cortex

Size

Catalog Number


C-12650 C-12651


C-14066

and the renal pelvis. The epithelial cells present in the distinct ducts and tubules differ in their morphology and physiological function. Consequently, HREpC are comprised of a heterogeneous population of epithelial cells. Important physiological processes like osmoregulation and excretion can be studied with HREpC in vitro.

Recommended media & reagents Epithelial Cell Growth Medium 2 (see page 68) DetachKit (see page 95) Cryo-SFM (see page 79) Renal

Related products Pellet (see page 92) Human Renal Cortical Epithelial Cells (see page 26) PromoFectin Transfection Reagent (see page 157) HREpC

Renale capsule Renal medulla Renal calices Renal pelvis Ureter

Fig. 5: Schematic structure of a kidney.

Fig. 6: HREpC culture in phase contrast.

26


https://www.promocell.com/shop Product Human Renal Epithelial Cells (HREpC)

Human Primary Cells

HREpC Pellet

Human Renal Cortical Epithelial Cells (HRCEpC) HRCEpC are isolated from the cortex of the human kidney and stain positive for cytokeratin. The renal cortex is the outer portion of the kidney between the renal capsule and the renal medulla. It contains the renal corpuscles, the proximal and distal convoluted tubules, and the

Size

Catalog Number


C-12665 C-12666


C-14067

cortical collecting ducts. Important physiological processes like osmoregulation and excretion can be studied with HRCEpC in vitro. Recommended media & reagents Renal Epithelial Cell Growth Medium 2 (see page 68) DetachKit (see page 95) Cryo-SFM (see page 79)

Related products Pellet (see page 92) Human Renal Epithelial Cells (see page 25) PromoFectin Transfection Reagent (see page 157) HRCEpC

https://www.promocell.com/shop Product Human Renal Cortical Epithelial Cells (HRCEpC) HRCEpC Pellet

Human Placental Epithelial Cells (HPlEpC) HPlEpC are isolated from the amniotic membrane and stain positive for cytokeratin. Placental Epithelial Cells are of fetal origin and form a single layer at the inner surface of the amnion. They protect the fetus against maternal leukocytes, neo-

Size

Catalog Number


C-12660 C-12662


C-14068

plastic cells, and pathogens. Cultured HPlEpC are suitable for physiological and pathological studies of amniotic functions. Recommended media & reagents Epithelial Cell Growth Medium (see page 69) DetachKit (see page 95) Cryo-SFM (see page 79)

Related products Villous Mesenchymal Fibroblasts (see page 29) HPlEpC Pellet (see page 92) PromoFectin Transfection Reagent (see page 157) Human

Placental

https://www.promocell.com/shop Product Human Placental Epithelial Cells (HPlEpC) HPIEpC Pellet

Size

Catalog Number


C-12395 C-12397


C-14069


27

1.6 Fibroblasts

Wound

healing research engineering research Regeneration studies Toxicological research Oncology Epithelial-mesenchymal interaction studies Tissue

Features CD90

positive

Fibroblasts are morphologically heterogeneous with diverse appearances depending on their location in the body and their activity. Injury of tissue displays a proliferative stimulus for fibroblasts and induces them to produce wound healing proteins. Therefore, fibroblasts are well suited for wound healing studies.

Fig. 1: NHDF culture in phase contrast.

Cryopreserved Available

at second passage from different tissues

like collagens, glycosaminoglycans, and glycoproteins. The composition of the extracellular matrix determines the physical properties of connective tissues.

Human Primary Cells

Applications

Fibroblasts are the most common cells in connective tissue. Their main function is to maintain the structural integrity of the connective tissue by continuously secreting extracellular matrix proteins

Normal Human Dermal Fibroblasts (NHDF) NHDF are isolated from the dermis of juvenile foreskin or adult skin from different locations like the face, the breasts, the abdomen, and the thighs*. They can be used for wound healing studies and dermatological research to investigate diseases like scleroderma, fibrosarcoma, fibrosis, xeroderma pigmentosum, and histiocytoma. Moreover, fibroblasts are important for cancer research, tissue regeneration, and tissue engineering studies.

Available Fibroblasts: NHDF (see below) HPF (see page 28) HAoAF (see page 28) HCF (see page 28) HUF (see page 29) HVMF (see page 29)

Fig. 2: NHDF culture stained for CD90. Nuclei counterstained with DAPI.

Recommended media & reagents for NHDF juvenile foreskin Fibroblast Growth Medium (see page 70) DetachKit (see page 95) Cryo-SFM (see page 79) Recommended media & reagents for NHDF adult donor Fibroblast Growth Medium 2 (see page 70) DetachKit (see page 95) Cryo-SFM (see page 79)

Related products Pellet (see page 92) NHDF adult Pellet (see page 92) Normal Human Epidermal Keratinocytes (see page 32) Normal Human Epidermal Melanocytes (see page 33) PromoFectin Transfection Reagent (see page 157) FGF-1 and FGF-2 (see page 179) NHDF

https://www.promocell.com/shop Product Normal Human Dermal Fibroblasts (NHDF) juvenile foreskin NHDF Pellet Normal Human Dermal Fibroblasts (NHDF) adult donor NHDF adult Pellet

Size

Catalog Number


C-12300 C-12350


C-14030


C-12302 C-12352


C-14031

* Normal Human Epidermal Keratinocytes (NHEK) and Normal Human Epidermal Melanocytes (NHEM) cultured in M2 medium from the same donor are available on request.

Human Primary Cells

28


Human Pulmonary Fibroblasts (HPF)

pulmonary hypertension can also be investigated with cultured HPF.

HPF are isolated from human lung tissue. They are often used for studying lung tissue repair, tissue remodeling after injury, or the response after pulmonary inflammation. Fibroblast disorders like pulmonary fibrosis or hypoxia-induced

Recommended media & reagents Fibroblast Growth Medium 2 (see page 70) DetachKit (see page 95) Cryo-SFM (see page 79)

Related products Pellet (see page 92) Human Pulmonary Microvascular Endothelial Cells (see page 21) Human Small Airway Epithelial Cells (see page 24) PromoFectin Transfection Reagent (see page 157) FGF-1 and FGF-2 (see page 179) HPF

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Human Aortic Adventitial Fibroblasts (HAoAF) HAoAF are isolated from human aortic adventitial tissue. HAoAF can be used for studies concerning vascular diseases e.g. neointimal formation, vascular remodeling, atherosclerosis, and hypertension. They are also often used for the examination of fibroblast disorders like

Size

Catalog Number


C-12360 C-12361


C-14035

fibrosis or other diseases linked to either imperfect or excessive accumulation of fibroblasts. Recommended media & reagents Fibroblast Growth Medium 2 (see page 70) DetachKit (see page 95) Cryo-SFM (see page 79)

Related products Pellet (see page 91) Human Aortic Endothelial Cells (see page 15) Human Aortic Smooth Muscle Cells (see page 37) PromoFectin Transfection Reagent (see page 157) FGF-1 and FGF-2 (see page 179) HAoAF

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Human Cardiac Fibroblasts (HCF) HCF are isolated from the ventricles of an adult heart. They play a central role in the maintenance of the extracellular matrix in the normal heart and the synthesis of growth factors and cytokines. Under pathophysiological conditions, cardiac fibroblasts are involved in scar formation

Size

Catalog Number


C-12380 C-12381


C-14037

after myocardial infarction, cardiac fibrosis, and cardiac hypertrophy. HCF can be used to study such processes in vitro. Recommended media & reagents Fibroblast Growth Medium 3 (see page 71) DetachKit (see page 95) Cryo-SFM (see page 79)

Related products Pellet (see page 91) Human Cardiac Myocytes (see page 11) Human Cardiac Microvascular Endothelial Cells (see page 20) PromoFectin Transfection Reagent (see page 157) FGF-1 and FGF-2 (see page 179) HCF

https://www.promocell.com/shop Product Human Cardiac Fibroblasts (HCF) HCF Pellet

Size

Catalog Number


C-12375 C-12377


C-14036


HUF are isolated from the uterine wall. During pregnancy, the uterine wall undergoes deep but reversible structural and biochemical changes. The uterine wall is the target of hormones and growth factors that act simultaneously and lead to extensive modifications of the extracellular matrix. This and other morphological and biochemical effects can be studied in vitro using HUF.


Related products Pellet (see page 92) Human Uterine Microvascular Endothelial Cell (see page 21) Human Uterine Smooth Muscle Cells (see page 41) PromoFectin Transfection Reagent (see page 157) FGF-1 and FGF-2 (see page 179) HUF

https://www.promocell.com/shop Product Human Uterine Fibroblasts (HUF) HUF Pellet

Human Villous Mesenchymal Fibroblasts (HVMF) HVMF are isolated from human villous placental tissue. They can be used as model systems in organ development research. In addition, they are useful for investigating the mechanism regulating human placental growth and function.

Size

Catalog Number


C-12385 C-12386


C-14038


Related products Pellet (see page 92) Human Placental Epithelial Cells (see page 26) PromoFectin Transfection Reagent (see page 157) FGF-1 and FGF-2 (see page 179) HVMF

https://www.promocell.com/shop Product Human Villous Mesenchymal Fibroblasts (HVMF) HVMF Pellet

Size

Catalog Number


C-12390 C-12391


C-14034

Human Primary Cells

Human Uterine Fibroblasts (HUF)

29

30


1.7 Follicle Dermal Papilla Cells Applications Hair

growth research testing Toxicological testing Drug

Features

Recommended media & reagents Follicle Dermal Papilla Cell Growth Medium (see page 72) DetachKit (see page 95) Cryo-SFM (see page 79) Related products Pellet (see page 91) PromoFectin Transfection Reagent (see page 157) HFDPC

Human Primary Cells

Alkaline

phosphatase positive at second passage Available from adult lateral scalp Available from donors with different hair colors Cryopreserved

Human Follicle Dermal Papilla Cells (HFDPC) are isolated from human dermis originating from the lateral scalp. Information on donor hair and skin color is available for each lot. HFDPC stain positive for alkaline phosphatase.

Fig. 1: HFDPC culture in phase contrast.

Sebaceous gland Hair shaft

Bulge

Matrix cells

Dermal papillae are embedded in a laminin and collagen IV rich extracellular matrix at the base of the hair follicles. They are essential for the induction and maintenance of hair growth. Since they have androgen receptors, they can be used for in vitro screening of androgen blocking reagents.

Dermal papilla

Fig. 2: Diagram showing the location of the dermal papilla in the hair follicle.

https://www.promocell.com/shop Product Human Follicle Dermal Papilla Cells (HFDPC) HFDPC Pellet

Size

Catalog Number


C-12071 C-12072


C-14005


31

1.8 Hepatocytes

Drug

testing

Toxicological

testing studies Liver physiology research Liver pathology research Oncology Metabolism

Features Cryopreserved

immediately after isolation 3 - 6 x 106 viable cells per cryovial Adherence > 50% Phase I reaction positive Phase II reaction positive Albumin positive Human Hepatocytes (HHpC) are isolated from the normal livers of single donors. After isolation, the cells are immediately cryopreserved and then intensively quality tested. Test criteria are cell viability, cell adherence efficiency, phase I metabolism, phase II metabolism, and albumin synthesis. The

cell adherence efficiency is at least 50%. The liver is the main metabolic organ in the body performing glycolysis, glycogenesis, amino acid metabolism, lipid metabolism, and ureagenesis. In addition, it also regulates the blood sugar level. Furthermore, hepatocytes eliminate toxic substances soon after absorption since the nutrition rich blood from the gastro-intestinal system first has to pass the liver. This detoxification consists of phase I and phase II reactions. The most important phase I enzyme system is the cytochrome P450 system. Consequently, cultured human hepatocytes are perfectly suited for in vitro metabolism and toxicity/detoxification studies prior to preclinical or clinical tests. Note: For culturing Hepatocytes, Collagen type I coated culture vessels are necessary. Under standard in vitro cell culture conditions, mature human hepatocytes usually do not survive for longer than 10-14 days and do not proliferate.

Recommended media & reagents Hepatocyte Growth Medium (see page 73) Hepatocyte Maintenance Medium (see page 73) DetachKit (see page 95) Cryo-SFM (see page 79) Related products Pellet (see page 91) PromoFectin-Hepatocyte Transfection Reagent (see page 158) HHpC

Fig 1. HHpC culture in phase contrast.

Count Living cells

Dead cells Cell Lab Quanta SC TM, Beckman Coulter, Inc.

37,5

25

12,5

Fig. 2: Flow cytometric analysis of PromoCell HHpC using 7-AAD staining. 77 % viable hepatocytes after thawing of the cryopreserved cells.

0 0

10

102

103

104

7-AAD

https://www.promocell.com/shop Product Human Hepatocytes (HHpC) HHpC Pellet

Size

Catalog Number

3 - 6 x 106 cryopreserved cells

C-12850


C-14085

Human Primary Cells

Applications

32


1.9 Keratinocytes Applications Wound

healing research development and differentiation studies Dermatological research Drug uptake studies Pharmaceutical testing Cosmetic and toxicological testing Oncology

Human Primary Cells

Epidermal

Features

body and minimizes moisture loss. Keratinocytes are also able to produce a variety of cytokines, growth factors, interleukins, and complement factors. Therefore keratinocytes are important for wound healing, inflammation, and immune response. Recommended media & reagents Keratinocyte Growth Medium 2 (see page 74) DetachKit (see page 95) Cryo-SFM (see page 79)

positive from juvenile foreskin or adult skin from different locations Available from single and pooled donors

Related products NHEK.f single donor Pellet (see page 92) NHEK.f pooled Pellet (see page 92) NHEK adult, single donor Pellet (see page 92) NHEK adult, pooled Pellet (see page 92) Normal Human Dermal Fibroblasts (see page 27) Normal Human Epidermal Melanocytes (see page 33) PromoFectin Transfection Reagent (see page 157)

Cytokeratin Available

Normal Human Epidermal Keratinocytes (NHEK) are available from single or pooled donors isolated from the epidermis of juvenile foreskin or adult skin from different locations like the face, the breasts, the abdomen, and the thighs*. They are the major cell type in the epidermis, making up about 90% of the cells. Epidermal keratinocytes originate in the stratum basale and move up through the layers of the epidermis. During this movement, they undergo gradual differentiation and morphology changes until they reach the stratum corneum, where they form a layer of nucleus-free, flat, and highly keratinized squamous cells. This layer forms an effective barrier to the entry of infectious agents into the

Stratum corneum Stratum granulosum Stratum spinosum

Stratum basale

Fig. 1: Structure of the epidermis.

Fig. 2: NHEK culture in phase contrast. Note the cobblestone shape typical for keratinocytes.

Fig. 3: NHEK culture stained for Cytokeratin. Nuclei counterstained with DAPI.

https://www.promocell.com/shop Product Normal Human Epidermal Keratinocytes (NHEK) juvenile foreskin, single donor NHEK.f single donor Pellet Normal Human Epidermal Keratinocytes (NHEK) juvenile foreskin, pooled NHEK.f pooled Pellet Normal Human Epidermal Keratinocytes (NHEK) adult, single donor NHEK adult, single donor Pellet Normal Human Epidermal Keratinocytes (NHEK) adult, pooled NHEK adult, pooled Pellet

Size

Catalog Number


C-12001 C-12002


C-14001


C-12005 C-12007


C-14003


C-12003 C-12004


C-14002


C-12006 C-12008


C-14004

* Normal Human Dermal Fibroblasts (NHDF) and Normal Human Epidermal Melanocytes (NHEM) cultured in M2 medium from the same donor are available on request.


33

1.10 Melanocytes

Dermatological

research research Melanogenesis studies Cosmetic and toxicological testing Skin compound screening Melanoma

Features Available

from juvenile foreskin or adult skin from different locations Available from light, moderate, and dark skin types Donor hair and eye color available on request Mel-5 positive Cultured in PMA-containing or PMA-free Medium Normal Human Epidermal Melanocytes (NHEM) are isolated from the epidermis of juvenile foreskin or adult skin from different locations including the face, the breasts, the abdomen, and the thighs*. They are located in the stratum basale, but branch out between the keratinocytes in suprabasal layers (see Fig. 1). About 5-10 % of the cells in the epidermis are melanocytes. The main purpose of melanocytes is the production of melanin, the protein responsible for the pigmentation of the skin, eyes, and hair. Melanin protects the cells in the skin and in deeper layers from the hazardous effects of UV radiation. It is produced and stored in melanosomes, which are located close to the melanocyte membrane, but it can also be transferred to neighboring keratinocytes. PromoCell melanocytes are isolated

using either the serum-free, PMA (Phorbol Myristate Acetate)-containing Melanocyte Growth Medium or the serumfree and PMA-free Melanocyte Growth Medium M2. Since PMA is a tumor promoting mitogen, it can interfere with experimental approaches. Therefore, we recommend using cells isolated in Melanocyte Growth Medium M2 and also using this medium for cultivation. Recommended media & reagents Melanocyte Growth Medium (see page 75)

Melanocyte

Growth Medium M2 (see page 75) DetachKit (see page 95) Cryo-SFM (see page 79) Related products Pellet (see page 92) NHEM.f M2 Pellet (see page 92) NHEM M2 adult Pellet (see page 92) Normal Human Dermal Fibroblasts (see page 27) Normal Human Epidermal Keratinocytes (see page 32) PromoFectin Transfection Reagent (see page 157) NHEM.f

Stratum corneum Stratum granulosum Stratum spinosum

Stratum basale

Melanosoma Keratinocytes Melanocyte

Fig. 1: Structure of the epidermis with the location of a melanocyte.

Fig. 2: NHEM cultured in Melanocyte Growth Medium in phase contrast.

Fig. 3: NHEM cultured in Melanocyte Growth Medium M2 in phase contrast.

https://www.promocell.com/shop Product Normal Human Epidermal Melanocytes (NHEM) juvenile foreskin NHEM.f Pellet Normal Human Epidermal Melanocytes (NHEM) juvenile foreskin, cultured in M2 Medium NHEM.f M2 Pellet Normal Human Epidermal Melanocytes (NHEM) adult donor, cultured in M2 Medium NHEM M2 adult Pellet

Size

Catalog Number


C-12400 C-12450


C-14040


C-12402 C-12452


C-14042


C-12403 C-12453


C-14043

* Normal Human Epidermal Keratinocytes (NHEK) and Normal Human Dermal Fibroblasts (NHDF) from the same donor are available on request.

Human Primary Cells

Applications

34


1.11 Osteoblasts Applications Bone

differentiation remodeling Bone physiology research Oncology Inflammatory studies Osteoporosis research Human Primary Cells

Bone

Features From

knee and hip femoral tissue at second passage Alkaline phosphatase positive Tested for mineralization Growth and Mineralization Media available Cryopreserved

Human Osteoblasts (HOB) are isolated from femoral trabecular bone tissue from the knee or hip joint region (lot specific source information is available on request). They stain positive for alkaline phosphatase and can be mineralized using PromoCell Mineralization Medium. Osteoblasts are of mesenchymal origin and are mainly responsible for bone formation and remodeling. In vivo, they produce a collagen type I rich extracellular matrix called the osteoid. During their development, osteoblasts undergo a maturation process called osteogenesis. When they become encased in the bone matrix, they stop proliferating and become osteocytes.

Recommended media & reagents Osteoblast Growth Medium (see page 76) Osteoblast Mineralization Medium (see page 76) DetachKit (see page 95) Cryo-SFM (see page 79)

Fig. 2: Alkaline phosphatase activity in HOB stained with BCIP/NBT.

Fig. 3: Mineralized HOB. Calcium deposits stained with Alizarin Red S.

Related products Pellet (see page 92) PromoFectin Transfection Reagent (see page 157) HOB

For more information please see www.promocell.com/knowledge-base

Fig. 1: HOB culture in phase contrast.

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Size

Catalog Number


C-12720 C-12760


C-14071


35

1.12 Preadipocytes

Proliferation/differentiation Physiological

studies and pathological

research metabolism research Obesity/diabetes studies Oncology (e.g. breast cancer) Fat

Features From

subcutaneous and visceral adipose tissue Available from different locations Cryopreserved at second passage Differentiation capacity to adipocytes Growth and Differentiation Media available Human White Preadipocytes (HWP) are isolated from adult subcutaneous or visceral adipose tissue from different locations (lot specific source information is available on request).

Adipose tissue is crucial in energy storage and metabolic homeostasis. An increase in adipose tissue results either from an enlargement of mature adipocytes or from the differentiation of adipocyte precursor cells (preadipocytes) into new mature adipocytes. These preadipocytes are already present in adipose tissues. They can proliferate throughout adult life and replace the cells that have differentiated into mature adipocytes. Since this differentiation process is controlled by a variety of growth factors and hormones, preadipocytes are suitable for the investigation of the physiological mechanisms controlling proliferation, differentiation, and function of adipose tissue.

Recommended media & reagents Preadipocyte Growth Medium (see page 77) Preadipocyte Differentiation Medium (see page 77) Adipocyte Nutrition Medium (see page 77) DetachKit (see page 95) Cryo-SFM (see page 79) Related products subcutaneous Pellet (see page 92) HWP visceral Pellet (see page 92) PromoFectin Transfection Reagent (see page 157) HWP

The PromoCell adipocyte cell culture system provides preadipocytes with optimized growth media and serum-free differentiation media.

Fig. 1: Left: Undifferentiated HWP (phase contrast). Middle: HWP after in vitro differentiation into adipocytes using the PromoCell Preadipocyte Differentiation Medium (phase contrast). Right: HWP after in vitro differentiation into adipocytes (Lipid droplets stained with Oil Red O).

https://www.promocell.com/shop Product Human White Preadipocytes (HWP) subcutaneous HWP subcutaneous Pellet Human White Preadipocytes (HWP) visceral HWP visceral Pellet

Size

Catalog Number


C-12730 C-12731


C-14072


C-12732 C-12733


C-14073

Human Primary Cells

Applications

36


1.13 Skeletal Muscle Cells Applications

Human Primary Cells

Gene therapy research Transplantation research Developmental/differentiation studies Tissue repair research Pharmacological testing Neuromuscular disease research

Features

liferate and fuse with damaged muscle fibers or with one another forming new myotubes. SkMC are optimal for in vitro muscle studies. They proliferate very well in the mitogen-rich PromoCell Skeletal Muscle Cell Growth Medium. Fusion to myotubes with typical multinucleated syncytia can be induced by using the PromoCell Skeletal Muscle Cell Differentiation Medium.

Recommended media & reagents Skeletal Muscle Cell Growth Medium (see page 78) Skeletal Muscle Cell Differentiation Medium (see page 78) DetachKit (see page 95) Cryo-SFM (see page 79) Related products Pellet (see page 92) PromoFectin Transfection Reagent (see page 157) SkMC

Sarcomeric myosin positive Available from different muscles Differentiation capacity to multi nucleate syncytia Growth and Differentiation Media available Human Skeletal Muscle Cells (SkMC) are isolated from different skeletal muscles (e.g. Musculus pectoralis major) from adult single donors (lot specific source information is available on request). They are positive for sarcomeric myosin and negative for smooth muscle specific α-actin. New skeletal muscle cells originate from quiescent satellite cells, which are located in the muscle fibers between the basal lamina and the sarcolemma. Quiescent satellite cells are activated by stimuli such as muscle damage. After activation, the cells, now called myoblasts, start to pro-

Fig. 1: SkMC culture in phase contrast.

Fig. 2: Differentiated SkMC culture in phase contrast. Arrows indicate multinucleated syncytia.


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Size

Catalog Number


C-12530 C-12580


C-14060


37

1.14 Smooth Muscle Cells

Organ

specific drug testing matrix interaction

Cell-extracellular

studies Physiological

and biochemical studies

Oncology Pathological

studies (e.g. hypertension) Atherosclerosis research

the walls of blood vessels and hollow organs like the bladder, the uterus, and the gastrointestinal tract and are responsible for involuntary movements like peristaltic contractions, which propel food along the gastrointestinal tract. PromoCell Smooth Muscle Cells are available from the aorta, the coronary artery, the pulmonary artery, the umbilical artery, the trachea, the bronchi, the bladder, the prostate, and the uterus.

Fig. 1: HTSMC culture in phase contrast. Human Primary Cells

Applications

Features muscle α-actin positive CD90 negative Routinely morphology tested Cryopreserved at second passage Smooth

Smooth muscle cells, which form nonstriated muscles, are structurally and functionally different from skeletal and cardiac muscle cells. They are located in

Available Smooth Muscle Cells: HAoSMC (see below) HCASMC (see page 38) HPASMC (see page 38) HUASMC (see page 39) HTSMC (see page 39) HBSMC (see page 40) HBdSMC (see page 40) HPrSMC (see page 41) HUtSMC (see page 41)

Fig. 2: HTSMC culture stained for smooth muscle α-actin. Nuclei counterstained with DAPI.

Human Aortic Smooth Muscle Cells (HAoSMC) HAoSMC are isolated from plaque-free regions of the human aorta (see Fig. 3) and stain positive for smooth muscle α-actin. The pulsatile pressure produced by the heart causes a cyclic distention of the aorta. The smooth muscle cells in the aortic wall subsequently contract it again. Changes in the arterial wall, associated with vascular diseases such as atherosclerosis and hypertension, strongly influence this process.

Tunica externa (Connective tissue)

Tunica media (Smooth muscle cells)

HAoSMC are suitable for studying the role of smooth muscle cells under normal or disease conditions in vitro.

Tunica intima (Endothelial cells)

Fig. 3: Diagram of a blood vessel. Recommended media & reagents Smooth Muscle Cell Growth Related products HAoSMC Pellet (see page 91) Medium 2 (see page 79) DetachKit (see page 95) Human Aortic Adventitial Fibroblasts Cryo-SFM (see page 79) (see page 28)

Human

Aortic Endothelial Cells (see page 15) PromoFectin Transfection Reagent (see page 157)

https://www.promocell.com/shop Product Human Aortic Smooth Muscle Cells (HAoSMC) HAoSMC Pellet

Size

Catalog Number


C-12533 C-12532


C-14053

38


Human Coronary Artery Smooth Muscle Cells (HCASMC) Left main coronary artery

Human Primary Cells

HCASMC are isolated from human coronary arteries (see Fig. 4) and stain positive for smooth muscle α-actin. Coronary arteries supply the heart with blood and obstruction in atherosclerosis causes myocardial infarction.

Circumflex coronary artery

HCASMC are qualified to study the function of smooth muscle cells in the development of atherosclerotic plaques or in other diseases like hypertension. They are also useful for stent-graft compatibility testing.

Right coronary artery

Left anterior descending coronary artery

Fig. 4: Location of the coronary arteries. Recommended media & reagents Smooth Muscle Cell Growth Medium 2 (see page 79) DetachKit (see page 95) Cryo-SFM (see page 79)

Related products Pellet (see page 91) Human Coronary Artery Endothelial Cells (see page 15) HCASMC

PromoFectin

Transfection Reagent (see page 157)

https://www.promocell.com/shop Product Human Coronary Artery Smooth Muscle Cells (HCASMC) HCASMC Pellet

Size

Catalog Number


C-12511 C-12512


C-14052

Human Pulmonary Artery Smooth Muscle Cells (HPASMC)

DetachKit

HPASMC are isolated from human pulmonary arteries and stain positive for smooth muscle α-actin. They are suited for studying the function of pulmonary arteries under normal and disease conditions (e.g. vasoconstriction induced by alveolar hypoxia). They are also useful for physiological studies of vascular smooth muscle cells.

Related products Pellet (see page 92) Human Pulmonary Artery Endothelial Cells (see page 16) PromoFectin Transfection Reagent (see page 157)

Recommended media & reagents Smooth Muscle Cell Growth Medium 2 (see page 79)

Cryo-SFM


HPASMC

Fig. 5: Top: HPASMC culture in phase contrast. Bottom: HPASMC culture stained for smooth muscle α-actin. Nucleus counterstained with DAPI. https://www.promocell.com/shop

Product Human Pulmonary Artery Smooth Muscle Cells (HPASMC) HPASMC Pellet

Size

Catalog Number


C-12521 C-12522


C-14051


HUASMC are isolated from the arteries of the umbilical cord and stain positive for smooth muscle α-actin. Physiological and biochemical processes of vascular diseases can be analyzed with HUASMC in culture.

Recommended media & reagents Smooth Muscle Cell Growth Medium 2 (see page 79) DetachKit (see page 95) Cryo-SFM (see page 79)

Related products HUASMC Pellet (see page 92) Human Umbilical Artery Endothelial Cells (see page 14) PromoFectin Transfection Reagent (see page 157)

https://www.promocell.com/shop Product Human Umbilical Artery Smooth Muscle Cells (HUASMC) HUASMC Pellet

Size

Catalog Number


C-12500 C-12550


C-14050

Human Tracheal Smooth Muscle Cells (HTSMC)

Paries membranaceus

HTSMC are isolated from the trachea (see Fig. 6) from single donors and stain positive for smooth muscle α-actin. They connect the endings of the tracheal cartilage by a contractile bridge called the paries membranaceus and consequently regulate the airway lumen. Under pathological conditions like asthma, the amount of smooth muscle cells in the trachea walls is increased leading to smooth muscle hypertrophy or hyperplasia. Hence, HTSMC are suitable for in vitro studies of asthma and other pulmonary diseases. Recommended media & reagents Smooth Muscle Cell Growth Medium 2 (see page 79) DetachKit (see page 95) Cryo-SFM (see page 79) Related products Pellet (see page 92)

HTSMC

Hyaline cartilage Mucosa

Fig. 6: Cross section through the trachea with c-shaped cartilage connected by the tracheal muscles. Human

Tracheal Epithelial Cells (see page 23) PromoFectin Transfection Reagent (see page 157)

https://www.promocell.com/shop Product Human Tracheal Smooth Muscle Cells (HTSMC) HTSMC Pellet

Size

Catalog Number


C-12565 C-12566


C-14056

Human Primary Cells

Human Umbilical Artery Smooth Muscle Cells (HUASMC)

39

40


Human Primary Cells

Human Bronchial Smooth Muscle Cells (HBSMC) HBSMC are isolated from the bronchi from single donors and stain positive for smooth muscle α-actin. They connect the endings of the bronchial cartilage and consequently regulate the airway lumen. Under pathological conditions like asthma, the amount of smooth muscle cells in the bronchi walls is increased

leading to smooth muscle hypertrophy or hyperplasia. Hence, HBSMC are suitable for in vitro studies of asthma and other pulmonary diseases. Recommended media & reagents Smooth Muscle Cell Growth Medium 2 (see page 79) DetachKit (see page 95) Cryo-SFM (see page 79)

Related products Pellet (see page 91) Human Bronchial Epithelial Cells (see page 24) PromoFectin Transfection Reagent (see page 157) HBSMC

https://www.promocell.com/shop Product Human Bronchial Smooth Muscle Cells (HBSMC) HBSMC Pellet

Size

Catalog Number


C-12561 C-12562


C-14055

Human Bladder Smooth Muscle Cells (HBdSMC) HBdSMC are isolated from the detrusor muscle from the urinary bladder (see Fig. 7) of single donors and stain positive for smooth muscle α-actin. The thick bladder wall contains smooth muscle cells arranged in muscle bundles, which empty the bladder by coordinated contraction. HBdSMC can be used for in vitro studies of physiological processes as well as the effect of drugs on the bladder. Recommended media & reagents Smooth Muscle Cell Growth Medium 2 (see page 79) DetachKit (see page 95) Cryo-SFM (see page 79)

Mucosa Submucosa

Detrusor Muscle

Adventitia

Fig. 7: Diagram of the bladder.

Related products Pellet (see page 91) Human Bladder Microvascular Endothelial Cells (see page 20) PromoFectin Transfection Reagent (see page 157) HBdSMC

https://www.promocell.com/shop Product Human Bladder Smooth Muscle Cells (HBdSMC) HBdSMC Pellet

Size

Catalog Number


C-12571 C-12572


C-14057


41

Human Prostate Smooth Muscle Cells (HPrSMC) HPrSMC are isolated from the prostate from single donors and stain positive for smooth muscle a-actin. The prostate produces secretions that get into the urethra through contraction of the smooth muscle cells. Prostate smooth muscle cells can be involved in prostitis and benign prostatic hyperplasia (BPH). HPrSMC can be used to study these prostate diseases or physiological processes in vitro. Recommended media & reagents Smooth Muscle Cell Growth Medium 2 (see page 79) DetachKit (see page 95) Cryo-SFM (see page 79)

Lateral lobes Capsule Human Primary Cells

Verumontanum Ejaculatory duct

Fig. 8: Diagram of the prostate. PromoFectin

Transfection Reagent (see page 157)

Related products HPrSMC Pellet (see page 92)

https://www.promocell.com/shop Product Human Prostate Smooth Muscle Cells (HPrSMC)

Size

Catalog Number


C-12573 C-12574


C-14058

HPrSMC Pellet

Human Uterine Smooth Muscle Cells (HUtSMC) HUtSMC are isolated from the myometrium (the middle layer of the uterine wall, see Fig. 9) and stain positive for smooth muscle α-actin. During pregnancy, hormones and growth factors cause an extensive hypertrophy of uterine smooth muscle cells. This and other morphological and biochemical effects on the uterus can be studied in vitro using HUtSMC. Recommended media & reagents Smooth Muscle Cell Growth Medium 2 (see page 79) DetachKit (see page 95) Cryo-SFM (see page 79)

Uterine tube Perimetrium (peritoneum) Myometrium (muscle)

Uterine cavity

Endometrium (epithelium) Cavity of cervix

Vagina Fig. 9: Diagram of the uterus. Human

Uterine Microvascular Endothelial Cells (see page 21)

Related products HUtSMC Pellet (see page 92) Fig. 9: Diagram of the uterus.

Product Human Uterine Smooth Muscle Cells (HUtSMC) HUtSMC Pellet

https://www.promocell.com/shop Size

Catalog Number


C-12575 C-12576


C-14059

42


Overview Human Primary Cells

Population Doublings

Page

Sarcomeric α-actinin+ Slow muscle myosin+

> 15

11

P2

Differentiation tested

> 10

12

5,000 - 10,000 cells per cm²

P1

vWF+ CD31+ Dil-Ac-LDL uptake+

> 15

13

C-22010 C-22011

5,000 - 10,000 cells per cm²

P1


> 15

13

C-12206

C-22011

5,000 - 10,000 cells per cm²

P1


> 15

13


C-12208

C-22011

5,000 - 10,000 cells per cm²

P1


> 15

13

Human Umbilical Vein Endothelial Cells (HUVEC) pre-screened

C-12205

C-22010 C-22011

5,000 - 10,000 cells per cm²

P1


> 15

13

Human Umbilical Artery Endothelial Cells (HUAEC)

C-12202

C-22010 C-22011

5,000 - 10,000 cells per cm²

P1


> 15

14


C-12271

C-22020 C-22022

5,000 - 10,000 cells per cm²

P2


> 15

15

Human Coronary Artery Endothelial Cells (HCAEC)

C-12221

C-22020 C-22022

5,000 - 10,000 cells per cm²

P2


> 15

15

Human Pulmonary Artery Endothelial Cells (HPAEC)

C-12241

C-22010 C-22011

5,000 - 10,000 cells per cm²

P2


> 15

16

Human Saphenous Vein Endothelial Cells (HSaVEC)

C-12231

C-22010 C-22011

5,000 - 10,000 cells per cm²

P2


> 15

16

Human Dermal Microvascular Endothelial Cells (HDMEC) juvenile foreskin

C-12210

C-22020 C-22022

10,000 - 20,000 cells per cm²

P2


> 15

17

Human Dermal Microvascular Endothelial Cells (HDMEC) adult donor

C-12212

C-22020 C-22022

10,000 - 20,000 cells per cm²

P2


> 15

17

Human Dermal Microvascular Endothelial Cells (HDMEC) pre-screened

C-12215

C-22020 C-22022

10,000 - 20,000 cells per cm²

P2


> 15

17

Human Dermal Blood Endothelial Cells (HDBEC) juvenile foreskin

C-12211

C-22020

10,000 - 20,000 cells per cm²

P2

PodoplaninProx-1CD31+

> 15

18

Human Dermal Blood Endothelial Cells (HDBEC) adult donor

C-12225

C-22020

10,000 - 20,000 cells per cm²

P2

PodoplaninProx-1CD31+

> 15

18

Human Dermal Lymphatic Endothelial Cells (HDLEC) juvenile foreskin

C-12216

C-22022

10,000 - 20,000 cells per cm²

P2

Podoplanin+ Prox-1+ CD31+

> 15

19

Human Dermal Lymphatic Endothelial Cells (HDLEC) adult donor

C-12217

C-22022

10,000 - 20,000 cells per cm²

P2

Podoplanin+ Prox-1+ CD31+

> 15

19

Human Bladder Microvascular Endothelial Cells (HBdMEC)

C-12291

C-22020 C-22022

10,000 - 20,000 cells per cm²

P2


> 15

20

Human Primary Cells

Product

Cat. No. Cryopr. Cells

Recommended Culture Media*

Plating Density

Passage after Thawing

Human Cardiac Myocytes (HCM)

C-12810

C-22070

10,000 - 15,000 cells per cm²

P2

Human Chondrocytes (HCH)

C-12710

C-27101

10,000 - 20,000 cells per cm²

Human Umbilical Vein Endothelial Cells (HUVEC) single donor

C-12200

C-22010 C-22011

Human Umbilical Vein Endothelial Cells (HUVEC) pooled

C-12203


* The media in this table are ready-to-use. For Media Kits see page 60.

Marker

43


Page


> 15

20

P2


> 15

21

10,000 - 20,000 cells per cm²

P2


> 15

21

C-22022

10,000 - 20,000 cells per cm²

P2


> 10

22

C-12620

C-21060

10,000 - 15,000 cells per cm²

P2

Cytokeratin+

> 15

23

Human Tracheal Epithelial Cells (HTEpC)

C-12644

C-21060

10,000 - 15,000 cells per cm²

P2

Cytokeratin+

> 15

23

Human Bronchial Epithelial Cells (HBEpC)

C-12640

C-21060

10,000 - 15,000 cells per cm²

P2

Cytokeratin+

> 15

24

Human Small Airway Epithelial Cells (HSAEpC)

C-12642

C-21070

10,000 - 15,000 cells per cm²

P2

Cytokeratin+

> 15

24

Human Mammary Epithelial Cells (HMEpC)

C-12650

C-21010

10,000 - 15,000 cells per cm²

P2

Cytokeratin+

> 15

25

Human Renal Epithelial Cells (HREpC)

C-12665

C-26001 C-26030

10,000 - 15,000 cells per cm²

P2

Cytokeratin+

> 15

25

Human Renal Cortical Epithelial Cells (HRCEpC)

C-12660

C-26001 C-26030

10,000 - 15,000 cells per cm²

P2

Cytokeratin+

> 15

26

Human Placental Epithelial Cells (HPlEpC)

C-12395

C-26040

10,000 - 15,000 cells per cm²

P2

Cytokeratin+

>5

26

Normal Human Dermal Fibroblasts (NHDF) juvenile foreskin

C-12300

C-23010

3,500 - 7,000 cells per cm²

P2

CD90+

> 15

27

Normal Human Dermal Fibroblasts (NHDF) adult donor

C-12302

C-23020

3,500 - 7,000 cells per cm²

P2

CD90+

> 15

27


C-12360

C-23020

3,500 - 7,000 cells per cm²

P2

CD90+

> 15

28

Human Aortic Adventitial Fibroblasts (HAoAF)

C-12380

C-23020

3,500 - 7,000 cells per cm²

P2

CD90+

> 10

28

Human Cardiac Fibroblasts (HCF)

C-12375

C-23025

3,500 - 7,000 cells per cm²

P2

CD90+

> 15

28

Human Uterine Fibroblasts (HUF)

C-12385

C-23020

3,500 - 7,000 cells per cm²

P2

CD90+

> 15

29

Human Villous Mesenchymal Fibroblasts (HVMF)

C-12390

C-23020

3,500 - 7,000 cells per cm²

P2

CD90+

> 15

29

Human Follicle Dermal Papilla Cells (HFDPC)

C-12071

C-26501

5,000 - 10,000 cells per cm²

P2

Alkaline phosphatase+

> 10

30

Human Hepatocytes (HHpC)

C-12850

C-25010 C-25020

150,000 cells per cm²

P0

Phase I reaction+ Phase II reaction+ Albumin+

n/a

31

Normal Human Epidermal Keratinocytes (NHEK) juvenile foreskin, single donor

C-12001

C-20011


P2

Cytokeratin+

> 15

32

Normal Human Epidermal Keratinocytes (NHEK) juvenile foreskin, pooled

C-12005

C-20011


P2

Cytokeratin+

> 15

32

Normal Human Epidermal Keratinocytes (NHEK) adult, single donor

C-12003

C-20011


P2

Cytokeratin+

> 15

32

Product



Plating Density


Human Cardiac Microvascular Endothelial Cells (HCMEC)

C-12285

C-22020 C-22022

10,000 - 20,000 cells per cm²

P2


C-12281

C-22020 C-22022

10,000 - 20,000 cells per cm²

Human Uterine Microvascular Endothelial Cells (HUtMEC)

C-12295

C-22020 C-22022

Human Brain Microvascular Endothelial Cells (HBMEC)

C-12287



Marker

Human Primary Cells


44



Page

Cytokeratin+

> 15

32

P2

Mel-5+

> 15

33

5,000 - 10,000 cells per cm²

P2

Mel-5+

> 15

33

C-24300

5,000 - 10,000 cells per cm²

P2

Mel-5+

> 15

33

C-12720

C-27001 C-27020

10,000 - 20,000 cells per cm²

P2

Alkaline phosphatase+ Mineralization tested

> 10

34

Human White Preadipocytes (HWP) subcutaneous

C-12730

C-27410 C-27436 C-27438


P2


> 10

35

Human White Preadipocytes (HWP) visceral

C-12732

C-27410 C-27436 C-27438


P2


> 10

35

Human Skeletal Muscle Cells (SkMC)

C-12530

C-23060 C-23061

3,500 - 7,000 cells per cm²

P2

Sarcomeric myosin+ Differentiation tested

> 15

36

Human Aortic Smooth Muscle Cells (HAoSMC)

C-12533

C-22062

7,500 - 10,000 cells per cm²

P2

Smooth muscle α-actin+

> 15

37

Human Coronary Artery Smooth Muscle Cells (HCASMC)

C-12511

C-22062

7,500 - 10,000 cells per cm²

P2


> 15

38


C-12521

C-22062

7,500 - 10,000 cells per cm²

P2


> 15

38


C-12500

C-22062

7,500 - 10,000 cells per cm²

P2


> 15

39


C-12565

C-22062

7,500 - 10,000 cells per cm²

P2


> 15

39

Human Bronchial Smooth Muscle Cells (HBSMC)

C-12561

C-22062

7,500 - 10,000 cells per cm²

P2


> 15

40

Human Bladder Smooth Muscle Cells (HBdSMC)

C-12571

C-22062

7,500 - 10,000 cells per cm²

P2


> 15

40

Human Prostate Smooth Muscle Cells (HPrSMC)

C-12573

C-22062

7,500 - 10,000 cells per cm²

P2


> 15

41

Human Uterine Smooth Muscle Cells (HUtSMC)

C-12575

C-22062

7,500 - 10,000 cells per cm²

P2


> 15

41

Human Primary Cells

Product



Normal Human Epidermal Keratinocytes (NHEK) adult, pooled

C-12006

C-20011


P2

Normal Human Epidermal Melanocytes (NHEM) juvenile foreskin

C-12400

C-24010

5,000 - 10,000 cells per cm²

Normal Human Epidermal Melanocytes (NHEM) juvenile foreskin, cultured in M2 Medium

C-12402

C-24300

Normal Human Epidermal Melanocytes (NHEM) adult donor, cultured in M2 Medium

C-12403

Human Osteoblasts (HOB)


Plating Density


Marker


45

Human Primary Cells by Tissue Type

Page

Human Mesenchymal Stem Cells from Adipose Tissue (hMSC-AT)

49

Human White Preadipocytes (HWP) subcutaneous

35

Human White Preadipocytes (HWP) visceral

35

n Bladder Human Bladder Microvascular Endothelial Cells (HBdMEC)

20

Human Bladder Smooth Muscle Cells (HBdSMC)

40

n Blood/Bone Marrow Human Mesenchymal Stem Cells from Bone Marrow (hMSC-BM)

49

Human Mononuclear Cells from Peripheral Blood (hMNC-PB) single donor, ultra-pure

56

Human Mononuclear Cells from Peripheral Blood (hMNC-PB) pooled, ultra-pure

56

Human Mononuclear Cells from Cord Blood (hMNC-CB) single donor, ultra-pure

57

Human Mononuclear Cells from Cord Blood (hMNC-CB) pooled, ultra-pure

57

Human CD34 Progenitor Cells from Cord Blood (hCD34+-CB) single donor

52

Human CD34+ Progenitor Cells from Cord Blood (hCD34+-CB) pooled

52

Human CD133 Progenitor Cells from Cord Blood (hCD133+-CB) single donor

53

Human CD133+ Progenitor Cells from Cord Blood (hCD133+-CB) pooled

53

Human CD14 Monocytes from Peripheral Blood (hMoCD14+-PB), single donor

54

Human CD14+ Monocytes from Peripheral Blood (hMoCD14+-PB), single donor, pre-screened

54

Human Monocytes from Peripheral Blood (hMo-PB), single donor Human Monocytes from Peripheral Blood (hMo-PB), single donor, pre-screened

+

+

Page

n Brain

n Adipose Tissue

+

Product

Human Brain Microvascular Endothelial Cells (HBMEC)

22

n Heart Human Cardiac Fibroblasts (HCF)

28

Human Cardiac Myocytes (HCM)

11

Human Cardiac Microvascular Endothelial Cells (HCMEC)

20

n Kidney Human Renal Epithelial Cells (HREpC)

25

Human Renal Cortical Epithelial Cells (HRCEpC)

26

n Large Vessels Human Aortic Adventitial Fibroblasts (HAoAF)

28


15

Human Aortic Smooth Muscle Cells (HAoSMC)

37

Human Coronary Artery Endothelial Cells (HCAEC)

15

Human Coronary Artery Smooth Muscle Cells (HCASMC)

38

Human Pulmonary Artery Endothelial Cells (HPAEC)

16


38

Human Umbilical Artery Endothelial Cells (HUAEC)

14


39

54


13

54


13


13


13

n Bone/Joint/Skeletal System Human Osteoblasts (HOB)

34

Human Chondrocytes (HCH)

12

Human Umbilical Vein Endothelial Cells (HUVEC) pre-screened

13

Human Skeletal Muscle Cells (SkMC)

36

Human Saphenous Vein Endothelial Cells (HSaVEC)

16

Human Primary Cells

Product

46


Product

Page

Human Primary Cells

31

n Mammary Tissue Human Mammary Epithelial Cells (HMEpC)

Page

n Respiratory System

n Liver Human Hepatocytes (HHpC)

Product

25

n Umbilical Cord Human Umbilical Artery Endothelial Cells (HUAEC)

14


39


13


13


13


13

Human Umbilical Vein Endothelial Cells (HUVEC) pre-screened Human Mesenchymal Stem Cells from Umbilical Cord Matrix (hMSC-UC)


23

Human Tracheal Epithelial Cells (HTEpC)

23


39

Human Bronchial Epithelial Cells (HBEpC)

24

Human Bronchial Smooth Muscle Cells (HBSMC)

40

Human Small Airway Epithelial Cells (HSAEpC)

24


28


21

n Skin Human Dermal Microvascular Endothelial Cells (HDMEC) juvenile foreskin

17

Human Dermal Microvascular Endothelial Cells (HDMEC) adult donor

17

13

Human Dermal Microvascular Endothelial Cells (HDMEC) pre-screened

17

49

Human Dermal Blood Endothelial Cells (HDBEC) juvenile foreskin

18

Human Dermal Blood Endothelial Cells (HDBEC) adult donor

18

Human Dermal Lymphatic Endothelial Cells (HDLEC) juvenile foreskin

19

Human Dermal Lymphatic Endothelial Cells (HDLEC) adult donor

19

Normal Human Dermal Fibroblasts (NHDF) juvenile foreskin

27

Normal Human Dermal Fibroblasts (NHDF) adult donor

27

Human Follicle Dermal Papilla Cells (HFDPC)

30

n Uterus Human Uterine Fibroblasts (HUF)

29

Human Uterine Microvascular Endothelial Cells (HUtMEC)

21

Human Uterine Smooth Muscle Cells (HUtSMC)

41

n Placenta Human Pericytes from Placenta (hPC-PL)

51

Normal Human Epidermal Keratinocytes (NHEK) juvenile foreskin, single donor

32

Human Placental Epithelial Cells (HPlEpC)

26

Normal Human Epidermal Keratinocytes (NHEK) juvenile foreskin, pooled

32

Human Villous Mesenchymal Fibroblasts (HVMF)

29

Normal Human Epidermal Keratinocytes (NHEK) adult, single donor

32

Normal Human Epidermal Keratinocytes (NHEK) adult, pooled

32

Normal Human Epidermal Melanocytes (NHEM) juvenile foreskin

33

Normal Human Epidermal Melanocytes (NHEM) juvenile foreskin, cultured in M2 Medium

33

Normal Human Epidermal Melanocytes (NHEM) adult donor, cultured in M2 Medium

33

n Prostate Human Prostate Smooth Muscle Cells (HPrSMC)

41

Human Stem and Blood Cells PromoCell offers a collection of adult stem cells, as well as differentiated blood cells from normal human bone marrow, umbilical cord tissue, placenta, adipose tissue, peripheral blood, and cord blood. All cells are obtained from approved donor programs at associated medical centers. High-quality media that simulate the in vivo environment as well as optimized differentiation media systems are also available for these cell types.

2


2

Human Stem and Blood Cells 2.1 Mesenchymal Stem Cells . . . . . . . . . . . . . . 49 2.2 Pericytes . . . . . . . . . . . . . . . . . . . . . . . . . . . 51 2.3 CD34+ Progenitor Cells . . . . . . . . . . . . . . . . 52 2.4 CD133+ Progenitor Cells . . . . . . . . . . . . . . . 53 2.5 Monocytes . . . . . . . . . . . . . . . . . . . . . . . . . 54 2.6 Mononuclear Cells . . . . . . . . . . . . . . . . . . . 56 Overview Human Stem and Blood Cells . . . . . . . . 58

2 Human Stem and Blood Cells

49

2.1 Mesenchymal Stem Cells

cell research medicine research Tissue engineering studies Developmental experiments

Human Mesenchymal Stem Cells from Bone Marrow (hMSC-BM)

Stem

Regenerative

Features adult stem cells for in vitro differentiation Routinely analyzed by flow cytometry Two Growth Media and five optmized Differentiation Media available

PromoCell hMSC-BM are harvested from normal human bone marrow. The cells are tested for their ability to differentiate in vitro into adipocytes, chondrocytes, osteoblasts, and neuronal cells.

Multipotent Tested

Mesenchymal Stem Cells (MSC), also termed Mesenchymal Stromal Cells, are multipotent cells that can differentiate into a variety of cell types and have the capacity for self renewal. MSC have been shown to differentiate in vitro or in vivo into adipocytes, chondrocytes, osteoblasts, myocytes, neurons, hepatocytes, and pancreatic islet cells. Optimized PromoCell media are available to support both the growth of MSC and their differentiation into several different lineages. Recent experiments suggest that differentiation capabilities into diverse cell types vary between MSC of different origin.

Human Mesenchymal Stem Cells from Umbilical Cord Matrix (hMSC-UC) PromoCell hMSC-UC are harvested from normal human umbilical cord matrix (Wharton’s jelly). The cells are tested for their ability to differentiate in vitro into adipocytes, chondrocytes, osteoblasts, and neuronal cells.

Human Mesenchymal Stem Cells from Adipose Tissue (hMSC-AT) PromoCell hMSC-AT are harvested from normal human adipose tissue. The cells are tested for their ability to differentiate in vitro into adipocytes, chondrocytes, osteoblasts, and neuronal cells.

Recommended media & reagents Stem Cell Growth Medium (see page 80) Mesenchymal Stem Cell Growth Medium DXF (see page 80) Mesenchymal Stem Cell Adipogenic Differentiation Medium (see page 80) Mesenchymal Stem Cell Chondrogenic Differentiation Medium (see page 80) Mesenchymal Stem Cell Chondrogenic Differentiation Medium w/o Inducers (see page 80) Mesenchymal Stem Cell Osteogenic Differentiation Medium (see page 80) Mesenchymal Stem Cell Neurogenic Differentiation Medium (see page 80) DetachKit (see page 95) Cell Dissociation Solution ACF (see page 96) Fibronectin Solution, human (see page 99) Mesenchymal

Related products MSC-Qualified

Fetal Calf Serum (see page 88) hMSC-BM Pellet (see page 92) hMSC-UC Pellet (see page 92) hMSC-AT Pellet (see page 92)


Fig. 1: Undifferentiated hMSC-BM cultured in MSC Growth Medium.

Fig. 2: Undifferentiated hMSC-BM cultured in MSC Growth Medium DXF.


Applications


50


Fig. 3.1

Fig. 3.1: hMSC-BM after in vitro differentiation into adipocytes using PromoCell Mesenchymal Stem Cell Adipogenic Differentiation Medium (lipid droplets stained with Oil Red O). Fig. 3.2: hMSC-BM spheroid after in vitro differentiation into cartilage using PromoCell Mesenchymal Stem Cell Chondrogenic Differentiation Medium (Alcian Blue staining).

Fig. 3.2

Fig. 3.3: hMSC-BM after in vitro differentiation into osteoblasts using PromoCell Mesenchymal StemCell Osteogenic Differentiation Medium (Alizarin Red S staining). Fig. 3.4: hMSC-BM after in vitro differentiation into neuronal cells using PromoCell Mesenchymal Stem Cell Neurogenic Differentiation Medium. Fig. 3.3

Fig. 3.4

For additional stem cell related products, please visit www.promokine.info/products/stem-cell-research

https://www.promocell.com/shop Product Human Mesenchymal Stem Cells from Bone Marrow (hMSC-BM) hMSC-BM Pellet Human Mesenchymal Stem Cells from Umbilical Cord Matrix (hMSC-UC) hMSC-UC Pellet Human Mesenchymal Stem Cells from Adipose Tissue (hMSC-AT) hMSC-AT Pellet

Size

Catalog Number


C-12974 C-12975


C-14090


C-12971 C-12972


C-14091


C-12977 C-12978


C-14092


51

2.2 Pericytes Applications Angiogenesis Regenerative

research medicine research

Oncology

blood vessel walls. They are important for angiogenesis, the structural integrity of the microvasculature, and blood flow regulation. However, they can also develop into malignant tumors called hemangiopericytomas.

Non-hematopoietic

multipotent mesenchymal-like cells Important in angiogenesis Routinely characterized by flow cytometry Isolated from human placenta Pericytes are multipotent mesenchymallike cells found in association with small

To date, pericytes have demonstrated the ability to differentiate into adipocytes, osteoblasts, fibroblasts, smooth muscle cells, and phagocytes (macrophages).

Fig. 4: Proliferating human pericytes (hPC-PL) from the placenta (phase contrast).

Recommended media & reagents Pericyte Growth Medium (see page 81) DetachKit (see page 95)

Related products hPC-PL Pellet (see page 92)

Count

CD146–


50

CD146+

25

Fig. 5: Flow cytometric analysis of PromoCell Pericytes. > 98% of the cells are CD146+.

0 100

101

102

103

104

CD146

https://www.promocell.com/shop Product Human Pericytes from Placenta (hPC-PL) hPC-PL Pellet

Size

Catalog Number


C-12980 C-12981


C-14095


Features

52


2.3 CD34+ Progenitor Cells Applications Stem

cell research basic research Hematological and immunological studies Vascular and angiogenesis research Oncology research

Features Multipotent

hematopoietic/ endothelial progenitors CD34+ cell purity > 95% Superior viability CD34 is a glycosylated transmembrane protein and represents a well-known

Recommended media & reagents Hematopoietic Progenitor Cell Expansion Medium DXF (see page 81) Hematopoietic Progenitor Medium (see page 81) Endothelial Progenitor Medium (see page 82) Related products (see page 179) SCF (see page 179) flt-3 ligand (see page 179) IL-3 (see page 179) IL-6 CC (see page 179) TPO (see page 179) CD34 antibody (see page 169) G-CSF

Count

CD34–



Transplantation

marker for primitive blood- and bone marrow-derived progenitor cells, especially for hematopoietic and endothelial stem cells. Although the biological functions of CD34 are largely unknown, recent data suggest that CD34 is involved in maintenance of the progenitor cells in a phenotypically undifferentiated state. PromoCell offers CD34+ Progenitor Cells from the cord blood of healthy donors. The CD34+ Progenitor Cells contain two main cellular subpopulations, hematopoietic and endothelial progenitor cells. Therefore, CD34+ Progenitor Cells are suitable for a series of studies, e.g. directed differentiation into more committed types of blood cells and endothelial lineages.

CD34+

50

25

Fig. 6: Flow cytometric analysis of PromoCell CD34+ Progenitor Cells. > 95% of the cells are CD34+.

0 0

10

102

103

104

CD34


Size

Catalog Number

Human CD34+ Progenitor Cells from Cord Blood (hCD34+-CB), single donor

100,000 cryopreserved cells

C-12921

Human CD34+ Progenitor Cells from Cord Blood (hCD34+-CB), pooled


C-12924


53

2.4 CD133+ Progenitor Cells

Hematology

research cell research Oncology Stem

Features progenitors, e.g. of hematopoietic lineages Available from cord blood CD133+ cell purity > 95% Superior viability Expansion and Differentiation Media available

CD133 (AC133, Prominin-1) is a glycoprotein with sub-cellular localization in plasma membrane protrusions. In humans, this protein was originally identified as an antigenic marker expressed on hematopoietic stem cells. In addition, CD133 is also expressed by neural and muscle progenitor cells.

Primitive

CD133 cells are isolated from fresh cord blood. These cells are highly proliferative primitive progenitors. They can give rise to mature terminally differentiated cells, e.g. hematopoietic lineages, by means of directed differentiation. PromoCell also provides suitable expansion and differentiation media. +

Recommended media & reagents Hematopoietic Progenitor Cell Expansion Medium DXF (see page 81) Cytokine Mix E for HPC Expansion Medium DXF (see page 81) Hematopoietic Progenitor Medium (see page 81) Related products G-CSF (see page 179) SCF (see page 179) flt-3 ligand (see page 179) IL-3 CC (see page 179) IL-6 CC (see page 179) TPO (see page 179) CD133 antibody (see page 169)

103

CD133+ cells 102

10

Fig. 7: Flow cytometric analysis of cord blood-derived CD133+ Progenitor Cells, demonstrating their high purity.

0 0

10

102


CD45

103

CD133


Size

Catalog Number



C-12927



C-12929


Applications

54


2.5 Monocytes Applications Dendritic

Cell Research

(Auto-)Immunity Infection

biology

Features Isolated


from fresh peripheral blood analyzed by flow cytometry CD14+-selected as well as “untouched“ Monocytes available

gradient-based method completely devoid of using any immunomagnetic beads. The advantage of these completely native Monocytes is that potential effects of bound magnetic particles (which can be ingested by this phagocytic cell type) are excluded.

Routinely

Monocytes (Mo) are immature phagocytic cells circulating in the blood. Acting as antigen-presenting immune cells, they can phagocytize and degrade microbes and particulate matter. Monocytes can differentiate into several different cell types. The most common applications for Monocytes are the in vitro differentiation into monocyte-derived dendritic cells (moDC), macrophages, and osteoclasts as well as their use as phagocytic immune cells in immunology and infection biology. CD14 belongs to the family of LPS receptor antigens and is strongly expressed on the majority of Monocytes. Thus, binding of immunomagnetic particles to this antigen is employed for purification of PromoCell CD14+-Monocytes (hMoCD14+-PB) from Mononuclear Cells (MNC). In contrast, the “untouched” Monocytes (hMo-PB) provided by PromoCell are enriched from MNC by a density

Human CD14+ Monocytes (hMoCD14+-PB) PromoCell Human CD14+ Monocytes are isolated from fresh peripheral blood. After isolation of the ultra-pure mononuclear cell fraction by proprietary methods, the CD14+ Monocytes are isolated by positive selection with > 95% purity. In addition, hMoCD14+-PB are available which are pre-screened for their differentiation capacity into dendritic cells.

Recommended media & reagents Mononuclear Cell Medium (see page 84) DC Generation Medium (see page 82) Related products hMoCD14+-PB single donor Pellet (see page 91) hMoCD14+-PB single donor pre-screened Pellet (see page 92) hMo-PB single donor Pellet (see page 92) hMo-PB single donor pre-screened Pellet (see page 92) PromoFectin-Macrophage Transfection Reagent (see page 158) G-CSF (see page 179) GM-CSF (see page 179) IL-4 CC (see page 179) TNF-alpha (see page 179) M-CSF (see page 179) sRANKL (see page 179) CD14 antibody (see page 169)

Human Monocytes (hMo-PB) PromoCell Human “untouched” Monocytes are isolated from fresh peripheral blood. After isolation of the ultra-pure mononuclear cell fraction by proprietary methods, the Monocyte fraction is further purified to > 70% by an advanced multi-gradient technique. In addition, hMo-PB are available which are pre-screened for their differentiation capacity into dendritic cells.

Fig. 8: Cluster formation during differentiation of Monocytes into moDC.


Fig. 9: Mature moDC cultured in PromoCell DC Generation Medium showing typical dendrite-like structures.


55


CD14+

CD14-

12,5

Fig. 10a: Flow cytometric analysis of day 8 mature moDCs generated in the PromoCell DC Generation Medium. More than 98% of the cells are negative for CD14.

0 0

101

102

103

104

103

104

CD14

Count Cell Lab Quanta SC TM, Beckman Coulter, Inc.

CD83+

CD83-

12,5

Fig. 10b: Flow cytometric analysis of day 8 mature moDCs generated in the PromoCell DC Generation Medium. The DC maturation marker CD83 is strongly expressed by > 98% of the cells.

0 0

101

102

CD83


Size

Catalog Number

10 x 106 cryopreserved cells

C-12909


C-14110


C-12912


C-14112


C-12910


C-14111

Human Monocytes from Peripheral Blood (hMo-PB), single donor, pre-screened


C-12913

hMo-PB single donor pre-screened Pellet


C-14113

Human CD14+ Monocytes from Peripheral Blood (hMoCD14+-PB), single donor hMoCD14+-PB single donor Pellet Human CD14+ Monocytes from Peripheral Blood (hMoCD14+-PB), single donor, pre-screened hMoCD14+-PB single donor pre-screened Pellet Human Monocytes from Peripheral Blood (hMo-PB), single donor hMo-PB single donor Pellet


Count

56


2.6 Mononuclear Cells Applications Stem

cell research basic research Hematological and immunological studies Oncology research Virology and infection biology

Features Routinely

analyzed by flow cytometry content < 5% Red blood cells depleted without lysis High purity and viability Available from single and pooled donors Granulocyte

matopoietic and endothelial progenitors, monocytes, and lymphocyte subpopulations (see Fig. 12).

Human Mononuclear Cells from Peripheral Blood (hMNC-PB) Adult peripheral blood contains large numbers of mature immune cells but also some progenitor cells. Typical application fields for hMNC-PB from single donors are immunology, oncology, hematology, virology, and infection biology. PromoCell hMNC-PB pooled are preferably used for several types of infectious virus tests, including neutralization assays. For HIV-related research, prior CD8+ T-cell depletion is recommended.

Size [µm]

Granulocytes Monocytes Cell Lab Quanta SC TM, Beckman Coulter, Inc.

16

14

Lymphocytes 12

Fig. 11: Flow cytometric analysis of Mononuclear Cells from Cord Blood (hMNC-CB). Granulocyte content 1%.

10 8 0 0

10

102

103

104

Side Scatter CD45 104

CD34+ cells 103



Transplantation

PromoCell Mononuclear Cells (MNC) represent the lymphocyte and monocyte fraction isolated from umbilical cord blood or peripheral blood. They are prepared by optimized low-density gradient centrifugation, which effectively reduces the granulocyte content (see Fig. 11). Red blood cells are gently but thoroughly depleted by proprietary techniques instead of osmotic lysis. This results in ultra-pure mononuclear cells with superior viability and unchanged biological function. In addition, the cells do not clump after thawing, allowing for easy handling and reproducible results. The MNC can be used directly for experiments or as a starting material for the isolation of several cell types, e.g., he-

102

10

Fig. 12: Flow cytometric determination of CD34+ cells within the PromoCell Mononuclear Cells from Cord Blood (hMNC-CB).

0 0

10

102

CD34

103

104


hMNC-CB are isolated from umbilical cord blood, which has a much higher content of primitive progenitor cells compared to adult peripheral blood. Recommended media & reagents Mononuclear Cell Medium (see page 84) Hematopoietic Progenitor Medium (see page 81)

Endothelial

Progenitor Medium (see page 82) Hematopoietic Progenitor Cell Expansion Medium DXF (see page 81) Cytokine Mix E for HPC Expansion Medium DXF (see page 81) Related products single donor Pellet (see page 91) hMNC-CB single donor Pellet (see page 91) hMNC-PB pooled Pellet (see page 91)

hMNC-CB

pooled Pellet (see page 91) Separation Medium 1077 (see page 84) GM-CSF (see page 179) IL-4 CC (see page 179) M-CSF (see page 179) TNF-alpha (see page 179) sRANKL (see page 179) Lymphocyte

hMNC-PB


Human Mononuclear Cells from Cord Blood (hMNC-CB)

57


Size

Catalog Number


C-12907


C-14098


C-12908


C-14099

Human Mononuclear Cells from Cord Blood (hMNC-CB), single donor, ultra-pure


C-12901

hMNC-CB single donor Pellet


C-14096

Human Mononuclear Cells from Cord Blood (hMNC-CB), pooled, ultra-pure


C-12904


C-14097

Human Mononuclear Cells from Peripheral Blood (hMNC-PB), single donor, ultra-pure hMNC-PB single donor Pellet Human Mononuclear Cells from Peripheral Blood (hMNC-PB), pooled, ultra-pure hMNC-PB pooled Pellet

hMNC-CB pooled Pellet

6

6

6

* These differentiation media are applicable for CFU-assays for hematopoietic progenitor cells (LTC-IC assays) or “Hill-assays” for endothelial progenitor cells (Hill et al. 2003, N Engl J Med 348: 593-600).

58


Overview Human Stem and Blood Cells


Product


Recommended Culture Media

Recommended Differentiation Media

Recommended Sera

Plating Density

Page

Human Mesenchymal Stem Cells from Bone Marrow (hMSC-BM)

C-12974

C-28010 C-28019

C-28011 C-28012 C-28013 C-28014 C-28015

C-37385 C-37386

4,000 cells per cm2

49

Human Mesenchymal Stem Cells from Umbilical Cord Matrix (hMSC-UC)

C-12971

C-28010 C-28019

C-28011 C-28012 C-28013 C-28014 C-28015

C-37385 C-37386

4,000 cells per cm2

49

Human Mesenchymal Stem Cells from Adipose Tissue (hMSC-AT)

C-12977

C-28010 C-28019

C-28011 C-28012 C-28013 C-28014 C-28015

C-37385 C-37386

4,000 cells per cm2

49

Human Pericytes from Placenta (hPC-PL)

C-12980

C-28040

C-37385 C-37386

4,000 cells per cm2

51


C-12921

C-28021 C-39890

C-28020* C-28025*

C-37387 C-37388

5,000 cells per ml

52


C-12924

C-28021 C-39890

C-28020* C-28025*

C-37387 C-37388

5,000 cells per ml

52


C-12927

C-28021 C-39890

C-28020*

C-37387 C-37388

5,000 cells per ml

53

Human CD133+ Progenitor Cells from Cord Blood (hCD133+CB), pooled

C-12929

C-28021 C-39890

C-28020*

C-37387 C-37388

5,000 cells per ml

53

Human CD14+ Monocytes from Peripheral Blood (hMoCD14+-PB), single donor

C-12909

C-28030

C-37387 C-37388

1 x106 cells per ml

54

Human CD14+ Monocytes from Peripheral Blood (hMoCD14+-PB), single donor, pre-screened

C-12912

C-28030

C-37387 C-37388

1 x106 cells per ml

54

Human Monocytes from Peripheral Blood (hMo-PB), single donor

C-12910

C-28030

C-37387 C-37388

1 x106 cells per ml

54

Human Monocytes from Peripheral Blood (hMo-PB), pre-screened

C-12913

C-28030

C-37387 C-37388

1 x106 cells per ml

54

Human Mononuclear Cells from Peripheral Blood (hMNCPB), single donor, ultra-pure

C-12907

C-28030

C-37387 C-37388

1 x106 cells per ml

56

Human Mononuclear Cells from Peripheral Blood (hMNCPB), pooled, ultra-pure

C-12908

C-28030

C-37387 C-37388

1 x106 cells per ml

56

Human Mononuclear Cells from Cord Blood (hMNC-CB), single donor, ultra-pure

C-12901

C-28030 C-28021 C-39890

C-28020* C-28025*

C-37387 C-37388

1 x106 cells per ml

57

Human Mononuclear Cells from Cord Blood (hMNC-CB), pooled, ultra-pure

C-12904

C-28030 C-28021 C-39890

C-28020* C-28025*

C-37387 C-37388

1 x106 cells per ml

57

C-28050

C-28050

* These differentiation media are applicable for CFU-assays for hematopoietic progenitor cells (LTC-IC assays) or “Hill-assays” for endothelial progenitor cells (Hill et al. 2003, N Engl J Med 348: 593-600).

Media and Sera High-quality media and sera are vital for successful cell culture. Therefore, PromoCell adheres to strict quality standards and produces media and sera only from raw materials from strictly approved sources. We also have invested a lot of effort in media optimization and have substituted serum by defined substances wherever possible. This ensures effective cell growth and avoids the unpredictable effects of indeterminate compounds.

3


3

Media and Sera 3.1

Media for Primary Cells . . . . . . . . . . . . . . 61

3.1.1

Myocyte Medium . . . . . . . . . . . . . . . . . . . . . 62

3.1.2

Chondrocyte Medium . . . . . . . . . . . . . . . . . . 62

3.1.3

Endothelial Cell Media . . . . . . . . . . . . . . . . . 63

3.1.4

Endothelial Cell Media MV . . . . . . . . . . . . . . 64

3.1.5

Epithelial Cell Media . . . . . . . . . . . . . . . . . . . 65

3.1.6

Fibroblast Media . . . . . . . . . . . . . . . . . . . . . . 70

3.1.7

Follicle Dermal Papilla Cell Medium . . . . . . . 72

3.1.8

Hepatocyte Media . . . . . . . . . . . . . . . . . . . . 73

3.1.9

Keratinocyte Medium . . . . . . . . . . . . . . . . . . 74

3.1.10 Melanocyte Media . . . . . . . . . . . . . . . . . . . . 75 3.1.11 Osteoblast Media . . . . . . . . . . . . . . . . . . . . . 76 3.1.12 Preadipocyte/Adipocyte Media . . . . . . . . . . . 77 3.1.13 Skeletal Muscle Cell Media . . . . . . . . . . . . . . 78 3.1.14 Smooth Muscle Cell Medium . . . . . . . . . . . . 79 3.1.15 Freezing Medium . . . . . . . . . . . . . . . . . . . . . 79

3.2

Media for Stem and Blood Cells . . . . . . . . . 80

3.2.1

Mesenchymal Stem Cell Media . . . . . . . . . . . 80

3.2.2

Pericyte Medium . . . . . . . . . . . . . . . . . . . . . 81

3.2.3

Hematopoietic Progenitor Cell Media . . . . . . 81

3.2.4

Endothelial Progenitor Medium . . . . . . . . . . 82

3.2.5

Dendritic Cell Generation Media . . . . . . . . . . 82

3.2.6

Macrophage Generation Media . . . . . . . . . . 83

3.2.7

Mononuclear Cell Medium . . . . . . . . . . . . . . 84

3.2.8

Blood Cell Separation Medium . . . . . . . . . . . 84

3.3

Classical Media . . . . . . . . . . . . . . . . . . . . . . 85

3.4

Insect Cell Culture Media . . . . . . . . . . . . . . 87

3.5 Sera . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87

3 Media and Sera

61

3.1 Media for Primary Cells convenience or as Medium Kit, which allows user control over the supplement type and concentration. Each medium lot is tested for its ability to support the growth of the intended primary cells. In addition, biochemical and

Medium (Ready-to-use)

in culture (including L-Glutamine and NaHCO3). The shelf life of the Basal Medium is 9 months (at 4 to 8°C), that of the SupplementMix is 12 months (at -20°C) from the date of manufacture. After adding the SupplementMix, the shelf life of the complete medium is 6 weeks (at 4 to 8°C).

The Medium (Ready-to-use) consists of a 500 ml bottle with Basal Medium and one vial with SupplementMix. Adding the SupplementMix to the Basal Medium results in the complete medium, which contains all the growth factors and supplements necessary for the propagation of the specific human primary cells

Medium Kit The Medium Kit consists of a 500 ml bottle with Basal Medium and a SupplementPack. The SupplementPack is a set of individual vials with pre-measured supplements. Adding all components of the SupplementPack to the Basal Medium results in the complete medium, which contains all the growth factors

sterility tests are performed*. Note: The media for primary cells do not contain antibiotics.

Media and Sera

The PromoCell media for primary cells are formulated for the optimal growth of specific types of primary human cells and a variety of primary animal cells. For most cell types, they are available as Medium (Ready-to-use) for maximal

and supplements necessary for the propagation of specific human primary cells in culture (including L-Glutamine and NaHCO3). Since all supplements can be added separately, the user has full control over the media formulation and can adapt it to his/her experimental requirements. The shelf life of the Basal Medium is 9 months (at 4 to 8°C), and that of the SupplementPack is 12 months (at -20°C) from the date of manufacture. After adding the supplements, the shelf life of the complete medium is 6 weeks (at 4 to 8°C). Basal Media (with or without phenol red) as well as SupplementMixes and SupplementPacks can also be purchased separately.

If you require special media modifications, we offer a Custom Media Service starting at 10 bottles per order. Please ask for details.

*Certificates of analysis are available on request.

62

3 Media and Sera

Low Serum 3.1.1 Myocyte Medium Myocyte Growth Medium Recommended for Cardiac Myocytes (see page 11)

Media and Sera

Human

The PromoCell Myocyte Growth Medium is a low serum (5% V/V) medium supplemented with well chosen hormones and growth factors. It was developed for the optimal expansion and maintenance of primary cardiac myocytes in culture. The

medium is optimized for primary human cells but can also be used for primary porcine cardiac myocytes. Related products Fetal Calf Serum (see page 87)

Final supplement concentrations (after addition to the medium) Fetal Calf Serum Epidermal Growth Factor (recombinant human)

0.05 ml / ml 0.5 ng / ml

Basic Fibroblast Growth Factor (recombinant human)

2 ng / ml

Insulin (recombinant human)

5 µg / ml


Size

Catalog Number

Myocyte Growth Medium (Ready-to-use)

500 ml

C-22070

Myocyte Growth Medium Kit

500 ml

C-22170

Myocyte Basal Medium

500 ml

C-22270

Myocyte Basal Medium, phenol red-free

500 ml

C-22275

Myocyte Growth Medium SupplementMix

for 500 ml

C-39275

Myocyte Growth Medium SupplementPack

for 500 ml

C-39270

3.1.2 Chondrocyte Medium Chondrocyte Growth Medium Recommended for Human

Chondrocytes (see page 12)

The PromoCell Chondrocyte Growth Medium is designed for the optimal cultivation of human chondrocytes. The fetal calf serum in the medium is strictly pretested for optimal performance.

The medium is optimized for primary human cells but can also be used for primary porcine and rat chondrocytes.

Final supplement concentration (after addition to the medium) Fetal Calf Serum

0.1 ml / ml


Size

Catalog Number

Chondrocyte Growth Medium (Ready-to-use)

500 ml

C-27101

Chondrocyte Basal Medium

500 ml

C-27111

Chondrocyte Basal Medium, phenol red-free

500 ml

C-27115

for 500 ml

C-39635

Chondrocyte Growth Medium SupplementMix

3 Media and Sera

63

Low Serum

Endothelial Cell Growth Medium Endothelial Cell Growth Medium 2 Recommended for Human

Umbilical Vein Endothelial Cells (see page 13) Human Umbilical Artery Endothelial Cells (see page 14) Human Pulmonary Artery Endothelial Cells (see page 16) Human Saphenous Vein Endothelial Cells (see page 16)

Final supplement concentrations (after addition to the medium) Fetal Calf Serum

Endothelial Cell Growth Medium and Endothelial Cell Growth Medium 2 are low-serum (2% V/V) media optimized for the cultivation of endothelial cells from large blood vessels. Endothelial Cell Growth Medium 2 lacks Endothelial Cell Growth Supplement (ECGS, bovine hypothalamic extract) but contains Insulin-like Growth Factor (Long R3 IGF) and Vascular Endothelial Growth Factor (VEGF). Generally, VEGF leads to higher endothelial cell proliferation in culture. But, because of its mul-

tiple effects on cell metabolism, it may also interfere with certain experimental setups. In these cases, we recommend the use of Endothelial Cell Growth Medium, which contains no VEGF. The media are optimized for primary human cells but can also be used for bovine, porcine, murine, caprine, rabbit, elephant, dog, and rat endothelial cells from large vessels as well as for various endothelial cell lines. Related products Fetal Calf Serum (see page 87)

Endothelial Cell Growth Medium

Endothelial Cell Growth Medium 2

0.02 ml / ml

0.02 ml / ml

0.004 ml / ml

—

0.1 ng / ml

5 ng / ml

1 ng / ml

10 ng / ml

Insulin-like Growth Factor (Long R3 IGF)

—

20 ng / ml

Vascular Endothelial Growth Factor 165 (recombinant human)

—

0.5 ng / ml

Endothelial Cell Growth Supplement Epidermal Growth Factor (recombinant human) Basic Fibroblast Growth Factor (recombinant human)

Ascorbic Acid Heparin Hydrocortisone

—

1 µg / ml

90 μg / ml

22.5 µg / ml

1 µg / ml

0.2 µg / ml


Size

Catalog Number

Endothelial Cell Growth Medium (Ready-to-use)

500 ml

C-22010

Endothelial Cell Growth Medium Kit

500 ml

C-22110

Endothelial Cell Basal Medium

500 ml

C-22210

Endothelial Cell Basal Medium, phenol red-free

500 ml

C-22215

Endothelial Cell Growth Medium SupplementMix

for 500 ml

C-39215

Endothelial Cell Growth Medium SupplementPack

for 500 ml

C-39210

Endothelial Cell Growth Medium 2 (Ready-to-use)

500 ml

C-22011

Endothelial Cell Growth Medium 2 Kit

500 ml

C-22111

Endothelial Cell Basal Medium 2

500 ml

C-22211

Endothelial Cell Basal Medium 2, phenol red-free

500 ml

C-22216

Endothelial Cell Growth Medium 2 SupplementMix

for 500 ml

C-39216

Endothelial Cell Growth Medium 2 SupplementPack

for 500 ml

C-39211

Media and Sera

3.1.3 Endothelial Cell Media

64

3 Media and Sera

Low Serum 3.1.4 Endothelial Cell Media MV Endothelial Cell Growth Medium MV Endothelial Cell Growth Medium MV2

Media and Sera

Recommended for Human

Dermal Microvascular Endothelial Cells (see page 17) Human Coronary Artery Endothelial Cells (see page 15) Human Aortic Endothelial Cells (see page 15) Human Pulmonary Microvascular Endothelial Cells (see page 21)

Final supplement concentrations (after addition to the medium) Fetal Calf Serum Endothelial Cell Growth Supplement Epidermal Growth Factor (recombinant human) Basic Fibroblast Growth Factor (recombinant human)

Human

Dermal Lymphatic Endothelial Cells (see page 19) Human Dermal Blood Endothelial Cells (see page 18) PromoCell Endothelial Cell Growth Medium MV and Endothelial Cell Growth Medium MV2 were developed for the in vitro cultivation of endothelial cells from microvascular vessels, the coronary artery, and the aorta. Endothelial Cell Growth Medium MV2 lacks the Endothelial Cell Growth Supplement (ECGS, bovine hypothalamic extract) but contains the basic Fibroblast Growth Factor, the Insulin-like Growth

Factor (Long R3 IGF-1) and the Vascular Endothelial Growth Factor (VEGF). Generally, VEGF leads to higher endothelial cell proliferation in culture. But, because of its multiple effects on the cell metabolism, it may also interfere with certain experimental setups. In these cases, we recommend the use of Endothelial Cell Growth Medium MV. The media are optimized for primary human cells but can also be used for bovine, porcine, murine, rat, and dog endothelial cells from microvascular vessels as well as for various endothelial cell lines. Related products Fetal Calf Serum (see page 87)

Endothelial Cell Growth Medium MV

Endothelial Cell Growth Medium MV2

0.05 ml / ml

0.05 ml / ml

0.004 ml / ml

—

10 ng / ml

5 ng / ml

—

10 ng / ml

Insulin-like Growth Factor (Long R3 IGF-1)

—

20 ng / ml

Vascular Endothelial Growth Factor 165 (recombinant human)

—

0.5 ng / ml

Ascorbic Acid

—

1 µg / ml

Heparin Hydrocortisone

90 μg / ml

—

1 µg / ml

0.2 µg / ml


Size

Catalog Number

Endothelial Cell Growth Medium MV (Ready-to-use)

500 ml

C-22020

Endothelial Cell Growth Medium MV Kit

500 ml

C-22120

Endothelial Cell Basal Medium MV

500 ml

C-22220

Endothelial Cell Basal Medium MV, phenol red-free

500 ml

C-22225

Endothelial Cell Growth Medium MV SupplementMix

for 500 ml

C-39225

Endothelial Cell Growth Medium MV SupplementPack

for 500 ml

C-39220

Endothelial Cell Growth Medium MV2 (Ready-to-use)

500 ml

C-22022

Endothelial Cell Growth Medium MV2 Kit

500 ml

C-22121

Endothelial Cell Basal Medium MV2

500 ml

C-22221

Endothelial Cell Basal Medium MV2, phenol red-free

500 ml

C-22226

Endothelial Cell Growth Medium MV2 SupplementMix

for 500 ml

C-39226

Endothelial Cell Growth Medium MV2 SupplementPack

for 500 ml

C-39221

3 Media and Sera

65

Serum-Free 3.1.5 Epithelial Cell Media


Nasal Epithelial Cells (see page 23) Human Bronchial Epithelial Cells (see page 24) Human Tracheal Epithelial Cells (see page 23)

The PromoCell Airway Epithelial Cell Growth Medium is a serum-free medium optimized for the cultivation of epithelial cells from large air passages. The medium is optimized for primary human cells but can also be used for murine, porcine, equine, and rat airway epithelial cells.

Media and Sera

Airway Epithelial Cell Growth Medium

Final supplement concentrations (after addition to the medium) Bovine Pituitary Extract Epidermal Growth Factor (recombinant human) Insulin (recombinant human)

0.004 ml / ml 10 ng / ml 5 μg / ml

Hydrocortisone

0.5 μg / ml

Epinephrine

0.5 μg / ml

Triiodo-L-thyronine

6.7 ng / ml

Transferrin, holo (human)

10 μg / ml

Retinoic Acid

0.1 ng / ml


Size

Catalog Number

Airway Epithelial Cell Growth Medium (Ready-to-use)

500 ml

C-21060

Airway Epithelial Cell Growth Medium Kit

500 ml

C-21160

Airway Epithelial Cell Basal Medium

500 ml

C-21260

Airway Epithelial Cell Basal Medium, phenol red-free

500 ml

C-21265

Airway Epithelial Cell Growth Medium SupplementMix

for 500 ml

C-39165

Airway Epithelial Cell Growth Medium SupplementPack

for 500 ml

C-39160

66

3 Media and Sera

Serum-Free Small Airway Epithelial Cell Growth Medium Recommended for Small Airway Epithelial Cells (see page 24)

Media and Sera

Human

PromoCell Small Airway Epithelial Cell Growth Medium is a serum-free medium developed for the cultivation of epithelial cells from the distal respiratory system. The optimal adjustment of growth factors in this medium ensures excellent in vitro growth.

The medium is optimized for primary human cells but can also be used for porcine small airway epithelial cells.

Final supplement concentrations (after addition to the medium) Bovine Pituitary Extract Epidermal Growth Factor (recombinant human) Insulin (recombinant human)

0.004 ml / ml 10 ng / ml 5 μg / ml

Hydrocortisone

0.5 μg / ml

Epinephrine

0.5 μg / ml

Triiodo-L-thyronine

6.7 ng / ml


10 μg / ml

Retinoic Acid

0.1 ng / ml

Bovine Serum Albumin-Fatty Acid Free (BSA-FAF)

2.5 mg / ml


Size

Catalog Number

Small Airway Epithelial Cell Growth Medium (Ready-to-use)

500 ml

C-21070

Small Airway Epithelial Cell Growth Medium Kit

500 ml

C-21170

Small Airway Epithelial Cell Basal Medium

500 ml

C-21270

Small Airway Epithelial Cell Basal Medium, phenol red-free

500 ml

C-21275

Small Airway Epithelial Cell Growth Medium SupplementMix

for 500 ml

C-39175

Small Airway Epithelial Cell Growth Medium SupplementPack

for 500 ml

C-39170

3 Media and Sera

67

Serum-Free Mammary Epithelial Cell Growth Medium Recommended for

The PromoCell Mammary Epithelial Cell Growth Medium is a serum-free medium developed for the cultivation of epithelial cells from the mammary gland.

The medium is optimized for primary human cells but can also be used for rat and murine mammary epithelial cells as well as mamma carcinoma cell lines.

Mammary Epithelial Cells (see page 25) Media and Sera

Human

Final supplement concentrations (after addition to the medium) Bovine Pituitary Extract Epidermal Growth Factor (recombinant human) Insulin (recombinant human) Hydrocortisone

0.004 ml / ml 10 ng / ml 5 μg / ml 0.5 μg / ml


Size

Catalog Number

Mammary Epithelial Cell Growth Medium (Ready-to-use)

500 ml

C-21010

Mammary Epithelial Cell Growth Medium Kit

500 ml

C-21110

Mammary Epithelial Cell Basal Medium

500 ml

C-21210

Mammary Epithelial Cell Basal Medium, phenol red-free

500 ml

C-21215

Mammary Epithelial Cell Growth Medium SupplementMix

for 500 ml

C-39115

Mammary Epithelial Cell Growth Medium SupplementPack

for 500 ml

C-39110

68

3 Media and Sera

Serum-Free/Low Serum Renal Epithelial Cell Growth Medium Renal Epithelial Cell Growth Medium 2

Media and Sera


Renal Epithelial Cells (see page 25) Human Renal Cortical Epithelial Cells (see page 26)

Final supplement concentrations (after addition to the medium) Fetal Calf Serum Epidermal Growth Factor (recombinant human) Insulin (recombinant human) Epinephrine Hydrocortisone

The PromoCell Renal Epithelial Cell Growth Medium is a serum-free medium for the cultivation of epithelial cells from the kidneys. The PromoCell Renal Epithelial Cell Growth Medium 2 is an optimized lowserum (5% V/V) medium with epinephrine. The combination of low serum and selected growth factors ensures successful growth in vitro and makes this medium the first choice for renal epithelial cells.

The media are optimized for primary human cells but can also be used for porcine renal epithelial cells. Related products Fetal Calf Serum (see page 87)

Renal Epithelial Cell Growth Medium

Renal Epithelial Cell Growth Medium 2

—

0.05 ml / ml

10 ng / ml

10 ng / ml

5 μg / ml

5 μg / ml

—

0.5 µg / ml

36 ng / ml

36 ng / ml


5 μg / ml

5 μg / ml

Triiodo-L-thyronine

4 pg / ml

4 pg / ml


Size

Catalog Number

Renal Epithelial Cell Growth Medium (Ready-to-use)

500 ml

C-26001

Renal Epithelial Cell Growth Medium Kit

500 ml

C-26011

Renal Epithelial Cell Basal Medium

500 ml

C-26021

Renal Epithelial Cell Basal Medium, phenol red-free

500 ml

C-26022

Renal Epithelial Cell Growth Medium SupplementMix

for 500 ml

C-39604

Renal Epithelial Cell Growth Medium SupplementPack

for 500 ml

C-39600

Renal Epithelial Cell Growth Medium 2 (Ready-to-use)

500 ml

C-26030

Renal Epithelial Cell Growth Medium 2 Kit

500 ml

C-26130

Renal Epithelial Cell Basal Medium 2

500 ml

C-26230

Renal Epithelial Cell Basal Medium 2, phenol red-free

500 ml

C-26235

Renal Epithelial Cell Growth Medium 2 SupplementMix

for 500 ml

C-39606

Renal Epithelial Cell Growth Medium 2 SupplementPack

for 500 ml

C-39605

3 Media and Sera

69

Low Serum


Placental Epithelial Cells (see page 26)

The PromoCell Placental Epithelial Cell Growth Medium is a low-serum (5% V/V) medium optimized for the cultivation of epithelial cells from the amniotic membrane of the placenta. The combination of low serum und selected growth factors ensures successful growth in vitro.

Related products Fetal Calf Serum (see page 87)

Media and Sera

Placental Epithelial Cell Growth Medium

Final supplement concentrations (after addition to the medium) Fetal Calf Serum Epidermal Growth Factor (recombinant human) Insulin (recombinant human)

0.05 ml / ml 10 ng / ml 5 μg / ml

Epinephrine

0.5 µg / ml

Hydrocortisone

36 ng / ml


5 μg / ml

Triiodo-L-thyronine

4 pg / ml


Size

Catalog Number

Placental Epithelial Cell Growth Medium (Ready-to-use)

500 ml

C-26040

Placental Epithelial Cell Growth Medium Kit

500 ml

C-26140

Placental Epithelial Cell Basal Medium

500 ml

C-26240

Placental Epithelial Cell Basal Medium, phenol red-free

500 ml

C-26245

Placental Epithelial Cell Growth Medium SupplementMix

for 500 ml

C-39612

Placental Epithelial Cell Growth Medium SupplementPack

for 500 ml

C-39610

70

3 Media and Sera

Serum-Free 3.1.6 Fibroblast Media Fibroblast Growth Medium Recommended for

The PromoCell Fibroblast Growth Medium is a serum-free medium optimized for the cultivation of neonatal and juvenile fibroblasts.

The medium is optimized for primary human cells but can also be used for rat, porcine, and bovine fibroblasts as well as fibroblast cell lines.

Human Dermal Fibroblasts, juvenile foreskin (see page 27)

Media and Sera

Normal

Final supplement concentrations (after addition to the medium) Basic Fibroblast Growth Factor (recombinant human)

1 ng / ml


5 μg / ml


Size

Catalog Number

Fibroblast Growth Medium (Ready-to-use)

500 ml

C-23010

Fibroblast Growth Medium Kit

500 ml

C-23110

Fibroblast Basal Medium

500 ml

C-23210

Fibroblast Basal Medium, phenol red-free

500 ml

C-23215

Fibroblast Growth Medium SupplementMix

for 500 ml

C-39315

Fibroblast Growth Medium SupplementPack

for 500 ml

C-39310

Low Serum Fibroblast Growth Medium 2 Recommended for Normal

Human Dermal Fibroblasts, adult donor (see page 27) Human Pulmonary Fibroblasts (see page 28) Human Aortic Adventitial Fibroblasts (see page 28) Human Uterine Fibroblasts (see page 29) Human Villious Mesenchymal Fibroblasts (see page 29)

PromoCell Fibroblast Growth Medium 2 is a low-serum (2% V/V) medium developed for adult human fibroblasts. The medium is optimized for primary human cells but can also be used for rat, porcine, and bovine fibroblasts as well as fibroblast cell lines.


0.02 ml / ml


1 ng / ml


5 μg / ml

Related products Calf Serum (see page 87)

Fetal

3 Media and Sera

71

Product

Size

Catalog Number

Fibroblast Growth Medium 2 (Ready-to-use)

500 ml

C-23020

Fibroblast Growth Medium 2 Kit

500 ml

C-23120

Fibroblast Basal Medium 2

500 ml

C-23220

Fibroblast Basal Medium 2, phenol red-free

500 ml

C-23225

Fibroblast Growth Medium 2 SupplementMix

for 500 ml

C-39325

Fibroblast Growth Medium 2 SupplementPack

for 500 ml

C-39320

Fibroblast Growth Medium 3 Recommended for Human

Cardiac Fibroblasts (see page 28)

PromoCell Fibroblast Growth Medium 3 has been developed for the in vitro cultivation of adult Human Cardiac Fibroblasts (HCF). The medium is optimized for primary human cells but can also be used for porcine cardiac fibroblasts.

Related products: calf serum (see page 87)

Fetal


0.1 ml / ml


1 ng / ml


5 μg / ml


Size

Catalog Number

Fibroblast Growth Medium 3 (Ready-to-use)

500 ml

C-23025

Fibroblast Growth Medium 3 Kit

500 ml

C-23130

Fibroblast Basal Medium 3

500 ml

C-23230

Fibroblast Basal Medium 3, phenol red-free

500 ml

C-23235

Fibroblast Growth Medium 3 SupplementMix

for 500 ml

C-39345

Fibroblast Growth Medium 3 SupplementPack

for 500 ml

C-39350

Media and Sera

https://www.promocell.com/shop

72

3 Media and Sera

Low Serum 3.1.7 Follicle Dermal Papilla Cell Medium Follicle Dermal Papilla Cell Growth Medium Recommended for Follicle Dermal Papilla Cells (see page 30)

Media and Sera

Human

The PromoCell Follicle Dermal Papilla Cell Growth Medium is a low-serum (4% V/V) medium. The combination of low serum and growth factors leads to excellent proliferation of human follicle dermal papilla cells in vitro.

Related products Fetal Calf Serum (see page 87)

Final supplement concentrations (after addition to the medium) Fetal Calf Serum Bovine Pituitary Extract

0.04 ml / ml 0.004 ml / ml


1 ng / ml


5 μg / ml


Size

Catalog Number

Follicle Dermal Papilla Cell Growth Medium (Ready-to-use)

500 ml

C-26501

Follicle Dermal Papilla Cell Growth Medium Kit

500 ml

C-26502

Follicle Dermal Papilla Cell Basal Medium

500 ml

C-26500

Follicle Dermal Papilla Cell Basal Medium, phenol red-free

500 ml

C-26505

Follicle Dermal Papilla Cell Growth Medium SupplementMix

for 500 ml

C-39625

Follicle Dermal Papilla Cell Growth Medium SupplementPack

for 500 ml

C-39620

3 Media and Sera

73

Serum-Free

Hepatocyte Growth Medium Hepatocyte Maintenance Medium Recommended for Human

Hepatocytes (see page 31)

The PromoCell Hepatocyte Growth Medium is a serum-free medium developed for the cultivation of human hepato-

Final supplement concentrations (after addition to the medium) Epidermal Growth Factor (recombinant human) Insulin (recombinant human)

cytes. It is supplemented with well chosen growth factors and hormones and suited for long-term cultivation.

The media are optimized for primary human hepatocytes but can also be used for porcine hepatocytes.

In situations where the media compounds of the Hepatocyte Growth Medium interfere with certain experimental setups, we recommend using the PromoCell Hepatocyte Maintenance Medium. This medium is developed for short-term cultivation and contains fewer supplements.

Note: For culturing Hepatocytes, Collagen type I coated culture vessels are necessary. Under standard in vitro cell culture conditions, mature human hepatocytes usually do not survive for longer than 10-14 days and do not proliferate.

Hepatocyte Growth Medium

Hepatocyte Maintenance Medium

10 ng / ml

—

5 μg / ml

6 ng / ml

Hydrocortisone

0.5 μg / ml

—


10 μg / ml

—

Ascorbic Acid Bovine Serum Albumin-Fatty Acid Free (BSA-FAF) Dexamethasone

250 µg / ml

—

3.75 mg / ml

—

—

400 ng / ml


Size

Catalog Number

Hepatocyte Growth Medium (Ready-to-use)

500 ml

C-25010

Hepatocyte Growth Medium Kit

500 ml

C-25110

Hepatocyte Basal Medium

500 ml

C-25210

Hepatocyte Basal Medium, phenol red-free

500 ml

C-25215

Hepatocyte Growth Medium SupplementMix

for 500 ml

C-39642

Hepatocyte Growth Medium SupplementPack

for 500 ml

C-39640

Hepatocyte Maintenance Medium (Ready-to-use)

500 ml

C-25020

Hepatocyte Maintenance Medium Kit

500 ml

C-25120

Hepatocyte Maintenance Medium SupplementMix

for 500 ml

C-39652

Hepatocyte Maintenance Medium SupplementPack

for 500 ml

C-39650

Media and Sera

3.1.8 Hepatocyte Media

74

3 Media and Sera

Serum-Free 3.1.9 Keratinocyte Medium Keratinocyte Growth Medium 2 Recommended for

Media and Sera

Normal

Human Epidermal Keratinocytes, juvenile foreskin, single donor (see page 32) Normal Human Epidermal Keratinocytes, juvenile foreskin, pooled (see page 32)

Normal

Human Epidermal Keratinocytes, adult, single donor (see page 32) Normal Human Epidermal Keratinocytes, adult, pooled (see page 32) PromoCell Keratinocyte Growth Medium 2 is a serum-free medium developed for the cultivation of epidermal keratino-

cytes without feeder cells. The medium is optimized for primary human cells but can also be used for horse and porcine keratinocytes, as well as mouse and rat keratinocytes after adapting the CaCl2 concentration. In addition, it is suited for rat corneal keratinocytes. Related products -Solution (see page 98) 2

CaCl

Final supplement concentrations (after addition to the medium) Bovine Pituitary Extract

0.004 ml / ml

Epidermal Growth Factor (recombinant human)

0.125 ng / ml


5 μg / ml

Hydrocortisone

0.33 μg / ml

Epinephrine

0.39 μg / ml

Transferrin, holo (human) CaCl2

10 μg / ml 0.06 mM


Size

Catalog Number

Keratinocyte Growth Medium 2 (Ready-to-use)

500 ml

C-20011

Keratinocyte Growth Medium 2 Kit

500 ml

C-20111

Keratinocyte Basal Medium 2

500 ml

C-20211

Keratinocyte Basal Medium 2, phenol red-free

500 ml

C-20216

Keratinocyte Growth Medium 2 SupplementMix

for 500 ml

C-39016

Keratinocyte Growth Medium 2 SupplementPack

for 500 ml

C-39011

3 Media and Sera

75

Serum-Free 3.1.10 Melanocyte Media

Recommended for Normal

Human Epidermal Melanocytes, juvenile foreskin (see page 33)

The PromoCell Melanocyte Growth Medium is a serum-free, PMA (Phorbol Myristate Acetate) containing medium optimized for the cultivation of human melanocytes. Since PMA is a tumor promoting mitogen, it may interfere with

some experimental approaches. In these cases, we recommend using Melanocyte Growth Medium M2. The medium is optimized for primary human cells but can also be used for horse, porcine, murine, and rat melanocytes. Media and Sera

Melanocyte Growth Medium

Final supplement concentrations (after addition to the medium) Bovine Pituitary Extract

0.004 ml / ml


1 ng / ml


5 μg / ml

Hydrocortisone

0.5 μg / ml

Phorbol Myristate Acetate

10 ng / ml


Size

Catalog Number

Melanocyte Growth Medium (Ready-to-use)

500 ml

C-24010

Melanocyte Growth Medium Kit

500 ml

C-24110

Melanocyte Basal Medium

500 ml

C-24210

Melanocyte Basal Medium, phenol red-free

500 ml

C-24215

Melanocyte Growth Medium SupplementMix

for 500 ml

C-39415

Melanocyte Growth Medium SupplementPack

for 500 ml

C-39410

Serum-Free Melanocyte Growth Medium M2 Recommended for Normal

Human Epidermal Melanocytes, juvenile foreskin, cultured in M2 Medium (see page 33)

Normal

Human Epidermal Melanocytes, adult donor, cultured in M2 Medium (see page 33) The unique PromoCell Melanocyte Growth Medium 2 is a serum-free and PMA (Phorbol Myristate Acetate) free

medium optimized for the cultivation of human melanocytes. It is optimized for primary human cells but can also be used for equine, porcine, murine, and rat melanocytes. It is also suitable for human pigment epithelial cells, neuronal cells and melanoma cells.

List of supplements and their concentrations: confidential https://www.promocell.com/shop Product

Size

Catalog Number

Melanocyte Growth Medium M2 (Ready-to-use)

500 ml

C-24300

Melanocyte Growth Medium M2 (Ready-to-use), phenol red-free

500 ml

C-24305

Melanocyte Basal Medium M2

500 ml

C-24400

Melanocyte Basal Medium M2, phenol red-free

500 ml

C-24405

for 500 ml

C-39420

Melanocyte Growth Medium M2 SupplementMix

76

3 Media and Sera

3.1.11 Osteoblast Media Osteoblast Growth Medium Recommended for

Media and Sera

Human

Osteoblasts (see page 34)

PromoCell Osteoblast Growth Medium was developed to establish and maintain human osteoblast cultures. The fetal calf serum in the medium is strictly pretested for optimal performance.

The media are optimized for primary human cells but can also be used for bovine, murine, and rat osteoblasts as well as osteoblast cell lines.

Final supplement concentration (after addition to the medium) Fetal Calf Serum

0.1 ml / ml


Size

Catalog Number

Osteoblast Growth Medium (Ready-to-use)

500 ml

C-27001

Osteoblast Basal Medium

500 ml

C-27010

Osteoblast Basal Medium, phenol red-free

500 ml

C-27015

for 500 ml

C-39615

Osteoblast Growth Medium SupplementMix

Osteoblast Mineralization Medium Recommended for Human

Osteoblasts (see page 34)

PromoCell Osteoblast Mineralization Medium was developed for the efficient induction of strong mineralization in human osteoblast cultures. Extensive extracellular calcium deposits can be ob-

served after 21 days, following an easy one-step protocol (see page 34, Fig. 3).

List of supplements and their concentrations: confidential https://www.promocell.com/shop Product Osteoblast Mineralization Medium

Size

Catalog Number

100 ml

C-27020

3 Media and Sera

77

Low Serum/Serum-Free 3.1.12 Preadipocyte/Adipocyte Media

Preadipocyte Differentiation Medium Adipocyte Nutrition Medium Recommended for Human

White Preadipocytes (see page 35)

The PromoCell Preadipocyte Growth Medium is a low-serum (5% V/V) medium optimized for the expansion of human preadipocytes. The Preadipocyte Differentiation Medium is a serum-free medium, which induces the differentiation of preadipocytes into mature adipocytes.

The media are optimized for primary human cells but can also be used for murine preadipocytes/adipocytes. Related products Calf Serum (see page 87)

Fetal

Media and Sera

Preadipocyte Growth Medium

The mature adipocytes can be cultivated using the Adipocyte Nutrition Medium.

Final supplement concentrations (after addition to the medium) Fetal Calf Serum Endothelial Cell Growth Supplement Epidermal Growth Factor (recombinant human) Hydrocortisone

Preadipocyte Growth Medium

Preadipocyte Differentiation Medium

Adipocyte Nutrition Medium

0.05 ml / ml

—

0.03 ml / ml

0.004 ml / ml

—

—

10 ng / ml

—

—

1 μg / ml

—

—

d-Biotin

—

8 μg / ml

8 μg / ml


—

0.5 μg / ml

0.5 μg / ml

Dexamethasone

—

400 ng / ml

400 ng / ml

IBMX

—

44 μg / ml

—

L-Thyroxine

—

9 ng / ml

—

Ciglitazone

—

3 μg / ml

—

90 μg / ml

—

—

Heparin


Size

Catalog Number

Preadipocyte Growth Medium (Ready-to-use)

500 ml

C-27410

Preadipocyte Growth Medium Kit

500 ml

C-27417

Preadipocyte Basal Medium

500 ml

C-27411

Preadipocyte Basal Medium, phenol red-free

500 ml

C-27415

Preadipocyte Growth Medium SupplementMix

for 500 ml

C-39425

Preadipocyte Growth Medium SupplementPack

for 500 ml

C-39427

Preadipocyte Differentiation Medium (Ready-to-use)

500 ml

C-27436

Preadipocyte Differentiation Medium Kit

500 ml

C-27437

Preadipocyte Differentiation Medium SupplementMix

for 500 ml

C-39436

Preadipocyte Differentiation Medium SupplementPack

for 500 ml

C-39437

Adipocyte Nutrition Medium (Ready-to-use)

500 ml

C-27438

Adipocyte Nutrition Medium Kit

500 ml

C-27439

Adipocyte Basal Medium

500 ml

C-27431

Adipocyte Basal Medium, phenol red-free

500 ml

C-27435

Adipocyte Nutrition Medium SupplementMix

for 500 ml

C-39438

Adipocyte Nutrition Medium SupplementPack

for 500 ml

C-39439

78

3 Media and Sera

Low Serum/Serum-Free 3.1.13 Skeletal Muscle Cell Media Skeletal Muscle Cell Growth Medium Skeletal Muscle Cell Differentiation Medium

Media and Sera


Skeletal Muscle Cells (see page 36)

The PromoCell Skeletal Muscle Cell Growth Medium is a low-serum (5% V/V) medium optimized for the expansion of human skeletal muscle cells.

The media are optimized for primary human cells but can also be used for rat, murine, and rabbit skeletal muscle cells.

The unique Skeletal Muscle Cell Differentiation Medium is a serum-free medium that induces the fusion of myoblasts to myotubes with typical multinucleated syncytia.

Fetal

Related products Calf Serum (see page 87)


Final supplement concentrations (after addition to the medium)

Skeletal Muscle Cell Growth Medium

Skeletal Muscle Cell Differentiation Medium

Fetal Calf Serum

0.05 ml / ml

—

Fetuin (bovine)

50 μg / ml

—

Epidermal Growth Factor (recombinant human)

10 ng / ml

—


1 ng / ml

—


10 μg / ml

10 μg / ml

Dexamethasone

0.4 μg / ml

—


Size

Catalog Number

Skeletal Muscle Cell Growth Medium (Ready-to-use)

500 ml

C-23060

Skeletal Muscle Cell Growth Medium Kit

500 ml

C-23160

Skeletal Muscle Cell Basal Medium

500 ml

C-23260

Skeletal Muscle Cell Basal Medium, phenol red-free

500 ml

C-23265

Skeletal Muscle Cell Growth Medium SupplementMix

for 500 ml

C-39365

Skeletal Muscle Cell Growth Medium SupplementPack

for 500 ml

C-39360

Skeletal Muscle Cell Differentiation Medium (Ready-to-use)

500 ml

C-23061

Skeletal Muscle Cell Differentiation Medium SupplementMix

for 500 ml

C-39366

3 Media and Sera

79

Low Serum

Smooth Muscle Cell Growth Medium 2 Recommended for Human

Umbilical Artery Smooth Muscle Cells (see page 39) Human Coronary Artery Smooth Muscle Cells (see page 38) Human Pulmonary Artery Smooth Muscle Cells (see page 38)

Human

Aortic Smooth Muscle Cells (see page 37) Human Bronchial Smooth Muscle Cells (see page 40) Human Tracheal Smooth Muscle Cells (see page 39) Human Bladder Smooth Muscle Cells (see page 40) Human Prostate Smooth Muscle Cells (see page 41) Human Uterine Smooth Muscle Cells (see page 41)

The PromoCell Smooth Muscle Cell Growth Medium 2 is a low-serum (5% V/V) medium developed to establish and maintain smooth muscle cell cultures. The medium is optimized for primary human cells but can also be used for bovine, porcine, murine, dog, and rat smooth muscle cells. Related products Fetal Calf Serum (see page 87)

Final supplement concentrations (after addition to the medium) Fetal Calf Serum Epidermal Growth Factor (recombinant human)

0.05 ml / ml 0.5 ng / ml


2 ng / ml


5 μg / ml


Size

Catalog Number

Smooth Muscle Cell Growth Medium 2 (Ready-to-use)

500 ml

C-22062

Smooth Muscle Cell Growth Medium 2 Kit

500 ml

C-22162

Smooth Muscle Cell Basal Medium 2

500 ml

C-22262

Smooth Muscle Cell Basal Medium 2, phenol red-free

500 ml

C-22267

Smooth Muscle Cell Growth Medium 2 SupplementMix

for 500 ml

C-39267

Smooth Muscle Cell Growth Medium 2 SupplementPack

for 500 ml

C-39262

3.1.15 Freezing Medium Recommended for Human

and Animal Primary Cells Cell Lines Induced Pluripotent Stem Cells Embryonic Stem Cells Established

Serum-Free The PromoCell Cryo-SFM is an animal component-free, chemically defined medium with DMSO, methylcellulose, and other cryoprotective components developed for the cryopreservation of human and animal primary cells. It ensures high

cell viability, excellent growth rates, and good attachment of the cells after thawing. The medium is the first choice for all types of primary cells and cell lines.

https://www.promocell.com/shop Product Cryo-SFM

Size

Catalog Number

30 ml 125 ml

C-29910 C-29912

Media and Sera

3.1.14 Smooth Muscle Cell Medium

80

3 Media and Sera

Media and Sera

3.2. Media for Stem and Blood Cells The PromoCell media for stem and blood cells are formulated for either the optimal growth of specific cell types or to support the differentiation into the respective mature cells. Each lot is tested for its ability to support growth or differentiation, respectively. In addition, biochemical and sterility tests are performed*. The Media (Ready-to-use) consist of a 100 ml, 250 ml, or 500 ml bottle of Basal Medium and a vial of SupplementMix. Adding the SupplementMix to the Basal Medium results in the complete medium.

Some Media are provided with additional Cytokine Mixes to achieve optimal performance. The shelf life of the Basal Medium is 9 months (at 4 to 8°C), and that of the SupplementMix is 12 months (at -20°C) from the date of manufacture. After adding the SupplementMix, the shelf life of the complete medium is 6 weeks (at 4 to 8°C). Note: The media for stem and blood cells do not contain antibiotics.

If you require special media modifications we offer a Custom Media Service starting at 10 bottles per order. Please ask for details.

3.2.1 Mesenchymal Stem Cell Media Mesenchymal Stem Cell Growth Medium Mesenchymal Stem Cell Growth Medium DXF Mesenchymal Stem Cell Differentiation Media Recommended for Human

Mesenchymal Stem Cells from Bone Marrow, Umbilical Cord Matrix, and Adipose Tissue (see page 49)

The PromoCell Mesenchymal Stem Cell Growth Medium supports the expansion of multipotent human Mesenchymal Stem Cells (MSC) without inducing early senescence and differentiation as observed in standard culture media. The Mesenchymal Stem Cell Growth Medium DXF allows for the robust proliferation of multipotent MSC in a chemically defined and xeno-free culture environment. It is devoid of all animal-derived components and substances of human origin with Human Serum Albumin as the only exception. Fibronectin-coated

plates are necessary in conjunction with this medium. In addition, PromoCell offers five media to efficiently induce differentiation of MSC into adipogenic, chondrogenic, osteogenic or neurogenic lineages, respectively (see also Fig. 3, page 50). Related products Cell Dissociation Solution ACF (see page 96) Fibronectin Solution, human (see page 99)


Size

Catalog Number

Mesenchymal Stem Cell Growth Medium (Ready-to-use)

500 ml

C-28010

Mesenchymal Stem Cell Growth Medium DXF (Ready-to-use)

500 ml

C-28019

Fibronectin Solution, human (1 mg/ml)

5 ml

C-43060

Mesenchymal Stem Cell Adipogenic Differentiation Medium (Ready-to-use)

100 ml

C-28011

Mesenchymal Stem Cell Chondrogenic Differentiation Medium (Ready-to-use)

100 ml

C-28012

Mesenchymal Stem Cell Chondrogenic Differentiation Medium w/o Inducers (Ready-to-use)

100 ml

C-28014

Mesenchymal Stem Cell Osteogenic Differentiation Medium (Ready-to-use)

100 ml

C-28013

Mesenchymal Stem Cell Neurogenic Differentiation Medium (Ready-to-use)

100 ml

C-28015

*Certificates of analysis are available on request.

3 Media and Sera

81

3.2.2 Pericyte Medium Pericyte Growth Medium Recommended for Human

Pericytes (see page 51)

PromoCell Pericyte Growth Medium has been developed and optimized for the in vitro cultivation of human pericytes from all tissues. Whereas in standard culture media peri-

cytes tend to grow slowly and induce early senescence, PromoCell Pericyte Growth Medium optimally supports cell proliferation and maintenance of multipotency in these cells.

List of supplements and their concentrations: confidential

Product Pericyte Growth Medium (Ready-to-use)

Size

Catalog Number

500 ml

C-28040

3.2.3 Hematopoietic Progenitor Cell Media Hematopoietic Progenitor Cell Expansion Medium DXF Hematopoietic Progenitor Medium Recommended for Human

Mononuclear Cells from Cord Blood (see page 57) Human CD34+ Progenitor Cells (see page 52) Human CD133+ Progenitor Cells (see page 53) Human Hematopoietic Progenitor Cells The PromoCell Hematopoietic Progenitor Cell (HPC) Expansion Medium DXF has been designed for the maximum expan-

sion of primitive HPCs in a fully defined, xeno-free culture environment. Due to the utilization of exclusively synthetic, recombinant or plant-sourced materials, this medium is chemically defined and free of all animal-derived components and substances of human origin with Human Serum Albumin as the only exception. Note: The medium must be supplemented with additional cytokines, e.g. Cytokine Mix E (see below). Individual cytokines can be found on our website www.promokine.info or page 179. Cytokine Mix E is a ready-to-use mixture of growth factors for the HPC Expansion Medium DXF. The animal component-free,

defined formula contains recombinant human TPO, SCF, flt-3 ligand and IL-3. The optimized formulation allows vigorous expansion of hematopoietic progenitor cells, while maintaining their primitive phenotype. Differentiation is largely inhibited. The PromoCell Hematopoietic Progenitor Medium has been developed for the expansion of hematopoietic progenitors by means of differentiation into more committed lineages of blood cells. Hence, this progenitor medium is suitable for CFU-based assays, e.g. long-term culture initiating cell (LTC-IC) assays.


List of supplements and their concentrations: confidential https://www.promocell.com/shop Product Hematopoietic Progenitor Cell Expansion Medium DXF Cytokine Mix E for HPC Expansion Medium DXF Hematopoietic Progenitor Medium (Ready-to-use)

Size

Catalog Number

500 ml

C-28021

1 ml (sufficient for 100 ml Medium) 5 ml (sufficient for 500 ml Medium)

C-39890 C-39891

100 ml

C-28020

Media and Sera


82

3 Media and Sera

3.2.4 Endothelial Progenitor Medium Endothelial Progenitor Medium Recommended for Mononuclear Cells (see page 56) Human CD34+ Progenitor Cells (see page 52) Human Endothelial Progenitor Cells Media and Sera

Human

The PromoCell Endothelial Progenitor Medium has been developed for the expansion of endothelial progenitors by means of differentiation into more committed lineages of endothelial cells. Hence, this progenitor medium is suitable for the differentiation of primitive progenitors into mature endothelial cells as well as for CFU-based assays, e.g. the

“Hill-Assay” for endothelial progenitor cells (Hill et al. 2003, N Engl J Med 348: 593-600), which is carried out on fibronectin-coated plates.

List of supplements and their concentrations: confidential https://www.promocell.com/shop Product Endothelial Progenitor Medium (Ready-to-use)

Size

Catalog Number

100 ml

C-28025

3.2.5 Dendritic Cell Generation Media DC Generation Medium DC Generation Medium DXF Monocyte Attachment Medium Recommended for Human

Monocytes, freshly isolated and cryopreserved (see page 54) Human Mononuclear Cells, freshly isolated The PromoCell Dendritic Cell (DC) Generation Media have been developed for the easy and efficient generation of immature as well as fully mature myeloid Dendritic Cells from peripheral blood monocytes.


PromoCell DC Generation Medium allows for the generation of DC from cryopreserved as well as freshly isolated cells. In the latter case, the Monocyte Attachment Medium is also needed. The DC Generation Medium DXF is a chemically defined and xeno-free formulation for use with freshly isolated cells. It is free of all animal derived components and substances of human origin with Human Serum Albumin as the only exception. The DC Base Media do not include cytokines and must therefore be supplemented according to the user’s individual needs.

The Monocyte Attachment Medium allows for the efficient adherence selection of Monocytes from freshly isolated Mononuclear Cells (MNC) while maintaining optimal cell health. Thus, the time-consuming and costly immunomagnetic purification of monocytes prior to dendritic cell generation is not necessary. Note: See our Instruction Manual and Application Note for details. Please visit www.promocell.com/application-notes.

3 Media and Sera

83

List of supplements and their concentrations: confidential

Product

Size

Catalog Number

DC Generation Medium (Ready-to-use)

250 ml

C-28050

DC Generation Medium DXF (Ready-to-use)

250 ml

C-28052

DC Base Medium

250 ml

C-28053

DC Base Medium DXF

250 ml

C-28054

Monocyte Attachment Medium (Ready-to-use)

250 ml

C-28051

3.2.6 Macrophage Generation Media M1-Macrophage Generation Medium DXF M2-Macrophage Generation Medium DXF Monocyte Attachment Medium Recommended for Human

Mononuclear Cells, freshly isolated Human Monocytes, freshly isolated The PromoCell Macrophage Generation Media were developed for easy and efficient generation of M1- or M2-polarized macrophages from peripheral blood monocytes. A detailed step-by-step protocol is available also, which allows optional user-customized macrophage activation.

Completely defined

In combination with the Monocyte Attachment Medium, freshly isolated peripheral blood mononuclear cells can be directly used as a starting material. The PromoCell M1-Macrophage Generation Medium DXF allows for the generation of M1-polarized macrophages from freshly isolated cells, whereas the M2-Macrophage Generation Medium DXF produces M2-polarized macrophages. The chemically defined and xeno-free media formulations are free of all animal derived components and substances of human origin with Human Serum Albumin as the only exception.

The Monocyte Attachment Medium allows for the efficient adherence selection of Monocytes from freshly isolated Mononuclear Cells (MNC) while maintaining optimal cell health. Thus, the time-consuming and costly immunomagnetic purification of monocytes prior to macrophage differentiation is not necessary. Note: See our Instruction Manual and Application Note for details. Please visit www.promocell.com/application-notes. Related products Detachment Solution DXF (see page 96)

Macrophage

The Macrophage Base Medium does not include cytokines and must therefore be supplemented according to the user’s individual needs.


Size

Catalog Number

M1-Macrophage Generation Medium DXF (Ready-to-use)

250 ml

C-28055

M2-Macrophage Generation Medium DXF (Ready-to-use)

250 ml

C-28056

Macrophage Base Medium DXF

250 ml

C-28057

Monocyte Attachment Medium (Ready-to-use)

250 ml

C-28051

Media and Sera


84

3 Media and Sera

3.2.7 Mononuclear Cell Medium Mononuclear Cell Medium

Human

Recommended for

The PromoCell Mononuclear Cell Medium is a very rich short-term maintenance medium which provides utmost cell viability.

Mononuclear Cells from Cord Blood (see page 57) Human Mononuclear Cells from Peripheral Blood (see page 56) Media and Sera

Human

Monocytes (see page 54)

48 hours before proceeding with your experiments. Related products Lymphocyte Separation Medium 1077 (see below)

It is suitable to maintain MNCs for up to

List of supplements and their concentrations: confidential https://www.promocell.com/shop Product Mononuclear Cell Medium (Ready-to-use)

Size

Catalog Number

500 ml

C-28030

3.2.8 Blood Cell Separation Medium Lymphocyte Separation Medium 1077 Recommended for Peripheral

blood cord blood Buffy coats Apheresis products Bone marrow Umbilical

The PromoCell Lymphocyte Separation Medium 1077 is a sterile, ready-to-use solution optimized for the separation of lymphocytes from whole blood or buffy coats without any loss of viability. It is based on Ficoll 400 and sodium diatrizoate and provides optimal physiological parameters. It has a density of 1,077 g/ml at +20°C.

https://www.promocell.com/shop Product Lymphocyte Separation Medium 1077

Size

Catalog Number

500 ml

C-44010

3 Media and Sera

85

3.3 Classical Media n DMEM Media Size

Catalog Number

DMEM, w/o L-Glutamine

500 ml

C-71110

DMEM, with L-Glutamine

500 ml

C-71111

DMEM, with 25 mM HEPES, w/o L-Glutamine

500 ml

C-71115

DMEM, with 25 mM HEPES, with L-Glutamine

500 ml

C-71116

DMEM, with stable Glutamine

500 ml

C-71120

DMEM, High Glucose (4.5 g/l), w/o L-Glutamine

500 ml

C-71210

DMEM, High Glucose (4.5 g/l), with L-Glutamine

500 ml

C-71212

DMEM, High Glucose (4.5 g/l), with stable Glutamine

500 ml

C-71220

DMEM, High Glucose (4.5 g/l), with stable Glutamine, with Na-Pyruvate

500 ml

C-71221

DMEM, High Glucose (4.5 g/l), with L-Glutamine, with Na-Pyruvate

500 ml

C-71222

DMEM, High Glucose (4.5 g/l), w/o L-Glutamine, with Na-Pyruvate

500 ml

C-71223

n DMEM/Ham´s F-12 Media Product


Catalog Number

DMEM/Ham´s F-12, w/o L-Glutamine

500 ml

C-71310

DMEM/Ham´s F-12, with L-Glutamine

500 ml

C-71311

DMEM/Ham´s F-12, with stable Glutamine

500 ml

C-71320

n Ham´s Media Product


Catalog Number

Ham´s F-10, w/o L-Glutamine

500 ml

C-72010

Ham´s F-10, with L-Glutamine

500 ml

C-72020

Ham´s F-12, w/o L-Glutamine

500 ml

C-72110

Ham´s F-12, with L-Glutamine

500 ml

C-72120

n Leibovitz L-15 Medium Product Leibovitz L-15, w/o L-Glutamine


Catalog Number

500 ml

C-73110

Media and Sera

Product


86

3 Media and Sera

n McCoy´s 5A Media

Media and Sera

Product


Catalog Number

McCoy’s 5A, w/o L-Glutamine

500 ml

C-73210

McCoy’s 5A, with L-Glutamine

500 ml

C-73211

McCoy´s 5A, with stable Glutamine

500 ml

C-73220

McCoy´s 5A, with stable Glutamine, phenol red-free

500 ml

C-73227

n MEM Alpha and MEM D-Valine Media Product


Catalog Number

MEM Alpha, w/o Nucleosides, w/o L-Glutamine

500 ml

C-75012

MEM Alpha, w/o Nucleosides, with L-Glutamine

500 ml

C-75013

MEM Alpha, with Nucleosides, with L-Glutamine

500 ml

C-75014

MEM D-Valine, w/o L-Glutamine

500 ml

C-75100

n MEM Earle Media Product


Catalog Number

MEM Earle, w/o L-Glutamine

500 ml

C-75210

MEM Earle, with L-Glutamine

500 ml

C-75211

MEM Earle, with stable Glutamine

500 ml

C-75220

n RPMI 1640 Media Product


Catalog Number

RPMI 1640, w/o L-Glutamine

500 ml

C-76010

RPMI 1640, with L-Glutamine

500 ml

C-76011

RPMI 1640, with 25 mM HEPES, w/o L-Glutamine

500 ml

C-76015

RPMI 1640, with 25 mM HEPES, with L-Glutamine

500 ml

C-76016

RPMI 1640, w/o L-Glutamine, phenol red-free

500 ml

C-76017

RPMI 1640, with stable Glutamine

500 ml

C-76020

3 Media and Sera

87

The medium requirements of insect cells are significantly different from vertebrate cell culture. For instance, the optimal pH range for most insect cells is more acidic

(pH 6.2 to 6.5). Also, the amino acid concentration needs to be higher than in vertebrate cell media.

Mitsuhashi/Maramorosh Insect Medium

Mitsuhashi and Maramorosh Insect Medium is well-suited for Mosquito cell culture and must be supplemented with FCS prior to use.

Recommended for Mosquito

Cells https://www.promocell.com/shop

Product Mitsuhashi/Maramorosh Insect Medium

Schneider’s Drosophila Medium Recommended for Dipteran

Size

Catalog Number

500 ml

C-77110

Schneider’s Drosophila Medium was originally developed for the culture of Drosophila cells but can also be used for the culture of other dipteran cell lines.

Cells https://www.promocell.com/shop

Product Schneider’s Drosophila Medium, w/o L-Glutamine

Size

Catalog Number

500 ml

C-77210

3.5 Sera Fetal Calf Serum Recommended for Primary

Human Cells (all types)

The PromoCell Fetal Calf Sera are high quality, pretested fetal calf serum batches. They are extensively tested for their ability to support in vitro expansion of human primary cells.

In addition, histochemical staining is performed to ensure that the cultured primary cells maintain their cell type specific marker expression.

https://www.promocell.com/shop Product Fetal Calf Serum (EU Quality) primary human cell culture tested

Size

Catalog Number

100 ml 500 ml

C-37355 C-37350

Media and Sera

3.4 Insect Cell Culture Media

88

3 Media and Sera

MSC-Qualified Fetal Calf Serum Recommended for Mesenchymal

Stem Cells (all types)

The PromoCell MSC-Qualified Fetal Calf Sera are high quality, pre-selected fetal calf serum batches. They are extensively tested for their ability to support in vitro expansion of human Mesenchymal Stem

Cells. In addition, histochemical staining is performed to ensure that the cultured stem cells maintain the ability to differentiate into diverse lineages.

Media and Sera

Fig. 1: Expansion and differentiation of Mesenchymal Stem Cells (MSC) using media with the PromoCell MSC-Qualified Fetal Calf Serum. Left: Undifferentiated hMSC-BM cultured for 4 passages (phase contrast). Right: The same cells after 14 days of adipogenic induction (Oil Red O staining of lipid droplets with Hematoxylin counterstain). https://www.promocell.com/shop Product MSC-Qualified Fetal Calf Serum

ESC-Qualified Fetal Calf Serum Recommended for Pluripotent

Embryonic Stem Cells Progenitor Cells Endothelial Progenitor Cells Hematopoietic

Size

Catalog Number

100 ml 500 ml

C-37386 C-37385

The PromoCell ESC-Qualified Fetal Calf Sera are high quality, pre-selected fetal calf serum batches. They are extensively tested for their ability to support undifferentiated, feeder-free culture of pluripotent murine embryonic stem cells in the presence of leukemia inhibitory

factor. The maintenance of pluripotency is assayed by immunostaining for basal pluripotency markers. In addition, this serum is also recommended for the culture of hematopoietic and endothelial progenitor cells.

Fig. 2: Growth morphology of murine embryonic stem cells (mESC). Left: mESC in standard Fetal Calf Serum. Note the large, flat, differentiated cells. Right: mESC after 5 passages in PromoCell ESC-Qualified Fetal Calf Serum. The cells are small and have a compact colony morphology typical for mESC. https://www.promocell.com/shop Product ESC-Qualified Fetal Calf Serum

Size

Catalog Number

100 ml 500 ml

C-37388 C-37387

Cell Pellets PromoCell Cell Pellets are an easily accessible source of RNA, DNA, or proteins from a wide range of human primary cell types without laborious cell cultivation work. This allows you to focus on your molecular biology experiments and maximize your valuable research time.

4


4

Cell Pellets 4.1

Cell Pellets . . . . . . . . . . . . . . . . . . . . . . . . . 91

4 Cell Pellets

91

4.1 Cell Pellets Indefinitely

stable at -20°C with established nucleic acid purification protocols

Compatible Gene

expression analysis characterization studies Northern blots Southern blots Western blots/2D gel electrophoresis Cell

Features Available

from all PromoCell cell types 1 million cells per pellet Stored in RNAlater®

PromoCell Cell Pellets are stored in RNAlater® - an aqueous protection solution that rapidly permeates the cells and stabilizes cellular DNA, RNA, and proteins and protects them against degradation. This makes the PromoCell Cell Pellets very useful for a wide range of research applications* like gene expression analysis, cell characterization studies, and protein characterization experiments.

Note: RNAlater® denatures proteins. Therefore, proteins will be suitable for e.g. Western blotting but not for applications that require native proteins. Each Cell Pellet is made from 1 million cells and is dissolved in 200 µl RNAlater®. The Cell Pellets are produced from released cell lots and have the same characteristics, i.e. expression profile or cell type specific antigens. On request the cells from the same lot are available for cultivation. The Cell Pellets can be stored indefinitely at -20°C. https://www.promocell.com/shop

Product

Product Description

HAoAF Pellet

Pellet from Human Aortic Adventitial Fibroblasts

C-14037

HAoEC Pellet

Pellet from Human Aortic Endothelial Cells

C-14023

HAoSMC Pellet

Pellet from Human Aortic Smooth Muscle Cells

C-14053

HBdMEC Pellet

Pellet from Human Bladder Microvascular Endothelial Cells

C-14026

HBdSMC Pellet

Pellet from Human Bladder Smooth Muscle Cells

C-14057

HBEpC Pellet

Pellet from Human Bronchial Epithelial Cells

HBMEC Pellet

Pellet from Human Brain Microvascular Endothelial Cells

HBSMC Pellet

Pellet from Human Bronchial Smooth Muscle Cells

C-14055

HCAEC Pellet

Pellet from Human Coronary Artery Endothelial Cells

C-14022

HCASMC Pellet

Pellet from Human Coronary Artery Smooth Muscle Cells

C-14052

HCF Pellet

Pellet from Human Cardiac Fibroblasts

C-14036

HCH Pellet

Pellet from Human Chondrocytes

C-14070

HCM Pellet

Pellet from Human Cardiac Myocytes

C-14080

HCMEC Pellet

Pellet from Human Cardiac Microvascular Endothelial Cells

C-14029

HDBEC adult Pellet

Pellet from Human Dermal Blood Endothelial Cells adult donor

C-14019

HDBEC Pellet

Pellet from Human Dermal Blood Endothelial Cells juvenile foreskin

C-14018

HDLEC adult Pellet

Pellet from Human Dermal Lymphatic Endothelial Cells adult donor

C-14021

HDLEC Pellet

Pellet from Human Dermal Lymphatic Endothelial Cells juvenile foreskin

C-14020

HDMEC adult Pellet

Pellet from Human Dermal Microvascular Endothelial Cells adult donor

C-14016

HDMEC Pellet

Pellet from Human Dermal Microvascular Endothelial Cells juvenile foreskin

C-14015

HDMEC pre-screened Pellet

Pellet from Human Dermal Microvascular Endothelial Cells pre-screened

C-14017

HFDPC Pellet

Pellet from Human Follicle Dermal Papilla Cells

C-14005

HHpC Pellet

Pellet from Human Hepatocytes

C-14085

HMEpC Pellet

Pellet from Human Mammary Epithelial Cells

C-14066

hMNC-CB pooled Pellet

Pellet from Human Mononuclear Cells from Cord Blood pooled, ultra-pure

C-14097

hMNC-CB single donor Pellet

Pellet from Human Mononuclear Cells from Cord Blood single donor, ultra-pure

C-14096

hMNC-PB single donor Pellet

Pellet from Human Mononuclear Cells from Peripheral Blood single donor, ultra-pure

C-14098

hMNC-PB pooled Pellet

Pellet from Human Mononuclear Cells from Peripheral Blood pooled, ultra-pure

C-14099

hMoCD14+-PB single donor Pellet

Pellet from Human CD14+ Monocytes from Peripheral Blood, single donor

C-14110

* More information on RNAlater® (FAQs, protocols) at www.invitrogen.com/site/us/en/home/brands/Product-Brand/rnalater.html

Catalog Number

C-14063 Limited Availability

C-14032

Cell Pellets

Applications

92

4 Cell Pellets

Cell Pellets


Product Description

Catalog Number

hMoCD14+-PB single donor pre-screened Pellet

Pellet from Human CD14+ Monocytes from Peripheral Blood, single donor pre-screened

C-14112

hMo-PB single donor Pellet

Pellet from Human Monocytes from Peripheral Blood, single donor

C-14111

hMo-PB single donor pre-screened Pellet

Pellet from Human Monocytes from Peripheral Blood, single donor pre-screened

C-14113

hMSC-AT Pellet

Pellet from Human Mesenchymal Stem Cells from Adipose Tissue

C-14092

hMSC-BM Pellet

Pellet from Human Mesenchymal Stem Cells from Bone Marrow

C-14090

hMSC-UC Pellet

Pellet from Human Mesenchymal Stem Cells from Umbilical Cord Matrix

C-14091

HNEpC Pellet

Pellet from Human Nasal Epithelial Cells

C-14062

HOB Pellet

Pellet from Human Osteoblasts

C-14071

HPAEC Pellet

Pellet from Human Pulmonary Artery Endothelial Cells

C-14024

HPASMC Pellet

Pellet from Human Pulmonary Artery Smooth Muscle Cells

C-14051

hPC-PL Pellet

Pellet from Human Pericytes from Placenta

C-14095

HPF Pellet

Pellet from Human Pulmonary Fibroblasts

C-14035

HPlEpC Pellet

Pellet from Human Placental Epithelial Cells

C-14069

HPMEC Pellet

Pellet from Human Pulmonary Microvascular Endothelial Cells

C-14027

HPrSMC Pellet

Pellet from Human Prostate Smooth Muscle Cells

C-14058

HRCEpC Pellet

Pellet from Human Renal Cortical Epithelial Cells

C-14068

HREpC Pellet

Pellet from Human Renal Epithelial Cells

C-14067

HSAEpC Pellet

Pellet from Human Small Airway Epithelial Cells

C-14065

HSaVEC Pellet

Pellet from Human Saphenous Vein Endothelial Cells

C-14025

HTEpC Pellet

Pellet from Human Tracheal Epithelial Cells

C-14064

HTSMC Pellet

Pellet from Human Tracheal Smooth Muscle Cells

C-14056

HUAEC Pellet

Pellet from Human Umbilical Artery Endothelial Cells

C-14013

HUASMC Pellet

Pellet from Human Umbilical Artery Smooth Muscle Cells

C-14050

HUF Pellet

Pellet from Human Uterine Fibroblasts

C-14038

HUtMEC Pellet

Pellet from Human Uterine Microvascular Endothelial Cells

C-14028

HUtSMC Pellet

Pellet from Human Uterine Smooth Muscle Cells

C-14059

HUVEC single donor Pellet

Pellet from Human Umbilical Vein Endothelial Cells, single donor

C-14010

HUVEC pooled Pellet

Pellet from Human Umbilical Vein Endothelial Cells, pooled

C-14011

HUVEC Growth Medium 2 single donor Pellet

Pellet from Human Umbilical Vein Endothelial Cells isolated in Growth Medium 2, single donor

C-14008

HUVEC Growth Medium 2 pooled Pellet

Pellet from Human Umbilical Vein Endothelial Cells isolated in Growth Medium 2, pooled

C-14009

HUVEC pre-screened Pellet

Pellet from Human Umbilical Vein Endothelial Cells pre-screened

C-14012

HVMF Pellet

Pellet from Human Villous Mesenchymal Fibroblasts

C-14034

HWP subcutaneous Pellet

Pellet from Human White Preadipocytes subcutaneous

C-14072

HWP visceral Pellet

Pellet from Human White Preadipocytes visceral

C-14073

NHDF adult Pellet

Pellet from Normal Human Dermal Fibroblasts adult donor

C-14031

NHDF Pellet

Pellet from Normal Human Dermal Fibroblasts juvenile foreskin

C-14030

NHEK adult, single donor Pellet

Pellet from Normal Human Epidermal Keratinocytes adult, single donor

C-14002

NHEK adult pooled Pellet

Pellet from Normal Human Epidermal Keratinocytes adult, pooled

C-14004

NHEK.f single donor Pellet

Pellet from Normal Human Epidermal Keratinocytes juvenile foreskin, single donor

C-14001

NHEK.f pooled Pellet

Pellet from Normal Human Epidermal Keratinocytes juvenile foreskin, pooled

C-14003

NHEM M2 adult Pellet

Pellet from Normal Human Epidermal Melanocytes adult donor, cultured in M2 Medium

C-14043

NHEM.f M2 Pellet

Pellet from Normal Human Epidermal Melanocytes juvenile foreskin, cultured in M2 Medium

C-14042

NHEM.f Pellet

Pellet from Normal Human Epidermal Melanocytes juvenile foreskin

C-14040

SkMC Pellet

Pellet from Human Skeletal Muscle Cells

C-14060

Reagents and Supplements PromoCell offers a broad range of reagents and supplements for cell culture. This includes enzyme mixtures for tissue and cell dissociation, buffers for daily cell culture work, antibiotics and antimycotics in different concentrations, components to supplement various culture media, such as cytokines and growth factors, and reagents needed for cell assays. All of these are intensively tested and meet our stringent quality criteria.

5


5

Reagents and Supplements 5.1

Detachment Reagents . . . . . . . . . . . . . . . . 95

5.2

Antibiotics and Antimycotics . . . . . . . . . . . . 97

5.3

Buffers and Salt Solutions . . . . . . . . . . . . . . 98

5.4 Supplements . . . . . . . . . . . . . . . . . . . . . . . 99

5 Reagents and Supplements

95

5.1 Detachment Reagents

Recommended for All

human primary cell types

The PromoCell DetachKit was developed for gentle and effective detachment of adherent primary human cells in culture. Each batch is tested on primary human cells.

The DetachKit is optimized for primary human cells but can also be used for primary animal cells and cell lines. The kit contains three components: HepesBSS (30 mM Hepes) Trypsin / EDTA Solution (0.04% / 0.03%) Trypsin Neutralizing Solution (TNS)

https://www.promocell.com/shop Product DetachKit

DetachKit 2 The PromoCell DetachKit 2 was developed for very gentle detachment of primary human cells in culture. It contains less trypsin and EDTA than the standard DetachKit and is suited for applications, which require very gentle cell handling.

Size

Catalog Number

30 ml 125 ml 250 ml

C-41200 C-41210 C-41220

The DetachKit 2 is optimized for primary human cells but can also be used for primary animal cells and cell lines. The kit contains three components: (30 mM Hepes) Trypsin / EDTA Solution (0.025% / 0.01%) Trypsin Neutralizing Solution (TNS) HepesBSS

https://www.promocell.com/shop Product DetachKit 2

Accutase-Solution Recommended for Primary

cells lines Tissue dissociation Cell

Size

Catalog Number

30 ml 125 ml 250 ml

C-41202 C-41212 C-41222

Accutase is an alternative cell detachment solution to trypsin and can also be used for tissue dissociation. It was developed for very gentle and effective detachment of adherent cells. The well balanced combination of proteolytic and collagenolytic activities ensures that sur-

face proteins and epitopes stay entirely intact. Accutase is therefore perfectly suited for applications, which require an unchanged cell surface. Each batch of the PromoCell AccutaseSolution is tested on primary human cells. https://www.promocell.com/shop

Product Accutase-Solution, primary human cell culture tested

Size

Catalog Number

100 ml

C-41310


DetachKit

96


Cell Dissociation Solution ACF Cell Dissociation Solution ACF is a proprietary non-enzymatic reagent for de-

tachment of adherent cell types from tissue culture plastic or glass surfaces. Due to its gentle yet effective action it presents a good animal component-free

alternative to trypsin and other enzymatic detachment solutions.



Cell Dissociation Solution ACF

Macrophage Detachment Solution DXF The Macrophage Detachment Solution DXF is a non-enzymatic reagent

Size

Catalog Number

100 ml

C-41320

especially optimized for the gentle detachment of strongly adherent macrophages.

https://www.promocell.com/shop Product Macrophage Detachment Solution DXF

Trypsin Solutions Trypsin is still the most widely employed enzyme used for cell detachment.

Size

Catalog Number

250 ml

C-41330

PromoCell´s Trypsin Solutions exhibit standardized enzyme activity and contain trypsin from extra-pure lots.


Size

Catalog Number

Trypsin / EDTA (0.04% / 0.03%)

30 ml 125 ml 250 ml

C-41000 C-41010 C-41020

Trypsin / EDTA 2 (0.025% / 0.01%)

30 ml 125 ml 250 ml

C-41002 C-41012 C-41022

Trypsin-EDTA PBS 1:250 (0.05% / 0.02%), w/o Ca++/Mg++

100 ml

C-41060

Trypsin-EDTA PBS 1:250, (0.5% / 0.2%), w/o Ca++/Mg++, 10x conc.

100 ml

C-41070

Trypsin Neutralizing Solution

https://www.promocell.com/shop Product Trypsin Neutralizing Solution (TNS) (0.05% Trypsin Inhibitor in 0.1% BSA)

Size

Catalog Number

30 ml 125 ml 250 ml

C-41100 C-41110 C-41120


97

5.2 Antibiotics and Antimycotics However, antibiotics may not only harm contaminating organisms, but can also unfavorably alter the physiology and lifespan of primary human cells. Therefore PromoCell does not recommend the routine use of antibiotics in cell

n Antibiotics and Antimycotics Product

culture as a general preventive measure. As a further application antibiotics may be utilized in genetic selection procedures, e.g. G-418.


Catalog Number

Pen/Strep Solution (10,000 IU/ml / 10 mg/ml)

100 ml

C-42010

Pen/Strep/Fungizone Solution (10,000 IU/ml / 10,000 μg/ml / 25 μg/ml)

100 ml

C-42020

Amphotericin B Solution (0.25 mg/ml)

100 ml

C-42040

Gentamicin Solution (5 mg/ml)

100 ml

C-42060

n Selection Antibiotics Product


Catalog Number

G-418 Sulphate Solution (50 mg/ml)

20 ml

C-42094

Hygromycin B Solution (50 mg/ml)

20 ml

C-42095


Antibiotics are used in order to prevent microbial contaminations in cell culture during initial isolation steps or to cure precious cultures exhibiting microbial contamination, e.g. with mycoplasma or yeast/fungi.

98


5.3 Buffers and Salt Solutions Buffers and Salt Solutions are frequently used in tissue culture, cell isolation procedures, and molecular biology.

For convenience, PromoCell offers ready-to-use Buffers and Salt Solutions. They are prepared from raw materials of

n Dulbecco´s Phosphate Buffered Saline


Product Dulbecco’s PBS

the highest quality using tissue culture grade water.


Catalog Number

500 ml

C-40230

Dulbecco’s PBS, w/o Ca /Mg

500 ml

C-40232

Dulbecco’s PBS, w/o Ca++/Mg++, 10x conc.

500 ml

C-40238

++

++

n Hanks´ Balanced Salt Solutions Product Hanks’ BSS


Catalog Number

500 ml

C-40310

++

Hanks’ BSS, w/o Ca /Mg

500 ml

C-40320

Hanks’ BSS, phenol red-free

500 ml

C-40370

Hanks’ BSS, w/o Ca /Mg , phenol red-free

500 ml

C-40390

++

++

++

n Endotoxin-free Buffers and Water Product


Catalog Number

PBS, endotoxin-free

500 ml

C-40240

PBS, w/o Ca /Mg , endotoxin-free

500 ml

C-40242

TC Water

500 ml

C-49998

++

++

n Other Solutions


Product

Size

Catalog Number

CaCl2-Solution (0.5 M)

2 ml

C-34006

HEPES Buffered Saline (30 mM HEPES)

30 ml 125 ml 250 ml

C-40000 C-40010 C-40020

HEPES Buffer (1 M)

100 ml

C-40030

Sodium bicarbonate Solution (7.5%)

100 ml

C-42340


99

5.4 Supplements

n Growth Supplements Size

Catalog Number

Endothelial Cell Growth Supplement* (bovine hypothalamic extract)

2 ml 10 x 2 ml

C-30160 C-30180

Endothelial Cell Growth Supplement/ Heparin* (bovine hypothalamic extract)

2 ml 10 x 2 ml

C-30120 C-30140

Bovine Pituitary Extract 15* (7.5 mg protein/ml)

2 ml 10 x 2 ml

C-30020 C-30040

Bovine Pituitary Extract 26* (13 mg protein/ml)

2 ml 10 x 2 ml

C-30021 C-30041

50 mg

C-52310

Human Transferrin APO, low iron content

1g

C-52322

Human Transferrin HOLO, iron saturated

1g

C-52332

Human Insulin, primary human cell culture tested

n Amino Acid Solutions Product


Catalog Number

L-Glutamine Solution

100 ml

C-42210

Stable Glutamine Solution

100 ml

C-42215

MEM Amino Acids Solution

100 ml

C-42320

MEM NEAA Solution

100 ml

C-42322

n Vitamin and Trace Element Solutions Product


Catalog Number

MEM Vitamin Solution

100 ml

C-42328

Trace Element Mix (1000x)

100 ml

C-42331

n Extracellular Matrix Proteins


Product

Size

Catalog Number

Fibronectin Solution, bovine (1 mg/ml)

5 ml

C-43050

Fibronectin Solution, human (1 mg/ml)

5 ml

C-43060

* Endothelial Cell Growth Supplement (ECGS) and Bovine Pituitary Extract (BPE) are popular growth supplements in primary cell culture. PromoCell ECGS and BPE are manufactured from raw material which is exclusively collected in New Zealand to ensure maximum BSE-safety.


Product


Cell Analysis The specific analysis of cellular processes and conditions has played an increasingly vital role in cell biology. Monitoring reporter gene expression in cells, determining cell viability and proliferation, testing for cytotoxicity, and distinguishing live and dead cells are important procedures in most cell biology labs. Measuring intracellular signaling and assaying distinct cell metabolic activities have also become routine. These applications all require reliable, sophisticated, and user-friendly research tools.

6


3 6

Cell Analysis 6.1 Cell Viability and Proliferation Kits . . . . . . 103 6.2 Live/Dead Cell Staining Kits . . . . . . . . . . . 105 6.3 Cytotoxicity Kits . . . . . . . . . . . . . . . . . . . . 106 6.4 Senescence Detection Kit . . . . . . . . . . . . . 106 6.5 Cell-Based Reporter Assays . . . . . . . . . . . 107 6.6 Cell Staining Reagents and Related Products . . . . . . . . . . . . . . . . . . . 109 6.7 Intracellular Indicators and Related Products . . . . . . . . . . . . . . . . . . . 115 6.8

Cell Signaling Assays . . . . . . . . . . . . . . . . 119

6.9

Cell Stress and Metabolism Kits . . . . . . . . 120

6.10 Cell Fractionation and Protein Extraction Kits . . . . . . . . . . . . . . . 122 6.11 Protein Size Markers and Protein Gel Stains . . . . . . . . . . . . . . . . . . . 123 6.12 Nucleic Acid Isolation Kits . . . . . . . . . . . . . 124 6.13 DNA Size Markers and DNA Gel Stains . . . 125

6 Cell Analysis

103

Applications

Cell viability and proliferation assays Growth factor/cytokine assays Optimization of cell culture conditions Cell number determination

Features Non-radioactive method Easy and fast procedure Reliable and reproducible results Appropriate for various multiwell formats and automation PromoKine offers several types of cell viability kits: The Fluorometric Cell Viability Kits I and II; the Colorimetric Cell Viability Kits I, II, III, and IV; and the Bioluminescent Cell Viability Kits I and II. Because all of the kits are designed for use with various multiwell formats, they are also suited for automation.

Fluorometric Cell Viability Kit I (Resazurin)

Fluorometric Cell Viability Kit II (Calcein)

The resazurin-based PromoKine Cell Viability Kit I provides a simple and rapid measurement of cell viability. Moreover, this single-step, homogenous cell proliferation assay is sensitive, safe, and costeffective. It only requires as few as 80 cells for reproducible results. The assay is based on the ability of viable cells to convert the non-fluorescent, blue redox dye Resazurin (also known as Alamar Blue) into the highly red fluorescent, pink dye resorufin.The fluorescent or colorimetric signal generated from the assay is proportional to the number of living cells in the sample. The kit reagents are nontoxic to cells and allow kinetic monitoring. While fluorometric resorufin quantitation is more sensitive, colorimetric measurement is also possible.

The PromoKine Fluorometric Cell Viability Kit II is intended for the reliable fluorometric determination of viable cell numbers. It utilizes the cell membranepermeable, non-fluorescent dye CalceinAM that is hydrolyzed by esterases into the cell membrane-impermeable greenfluorescent Calcein (Fig. 1). Since esterases are only active in viable cells, the amount of Calcein-AM hydrolyzed by cellular esterases is directly proportional to the number of viable cells in culture. The kit has a detection range from less than 50 to more than 25,000 cells per well.

The Colorimetric Cell Viability Kits I and II allow for easy and reliable colorimetric determination of viable cell numbers with excellent sensitivity and linearity. The kits utilize the tetrazolium salts WST-8 (Kit I) and WST-1 (Kit II) that are reduced to water-soluble, orange formazan dyes by dehydrogenases present in viable cells (see Fig. 2). The absorbance of the formazan dye is proportional to the number of metabolically active cells and can be measured directly from a 96well plate without additional processing.

Calcein-AM R: acetoxymethyl Esterase

Viable cell

Colorimetric Cell Viability Kits I (WST-8) and II (WST-1)

Calcein

Colorimetric Cell Viability Kits III (XTT) and IV (MTT) Fig. 1: Assay mechanism of Calcein-AM conversion by intracellular esterases.

WST-8 Formazan orange dye

WST-8 slightly yellow

1-Methoxy PMS

Viable cell

NADH, NADPH Dehydrogenase NAD, NADP

Fig. 2: Assay mechanism of WST-8 conversion by dehydrogenases in viable cells.

The PromoKine Colorimetric Cell Viability Kits III and IV were developed for determining the number of viable cells in culture. They are based on the reduction of the tetrazolium salts XTT or MTT into colored formazan compounds by mitochondrial enzymes. Since these enzymes are inactivated shortly after cell death, this is a reliable method for the detection of viable cells. The absorbance of the formazan product can be measured directly from a 96-well plate without additional processing.

Cell Analysis

6.1 Cell Viability and Proliferation Kits

Cell Analysis

104

6 Cell Analysis

Bioluminescent Cell Viability Kit I (ATP)

Bioluminescent Cell Viability Kit II (ADP/ATP)

The Promokine Bioluminescent Cell Viability Kit I employs bioluminescence to measure the ATP level as a marker for cell viability in mammalian cells. It utilizes luciferase to catalyze the ATP dependent, light-producing oxidation of luciferin. This light formation can be detected using a luminometer or beta counter. The assay can be performed directly in culture plates requiring no harvesting, washing, or sample preparation and can be fully automated for high throughput (10 seconds/sample). It is highly sensitive, detecting 10-100 mammalian cells per well. It also offers applications besides cell viability testing, e.g., detecting low level bacterial contamination in blood, milk, urine, soil, and sludge.

The ADP/ATP ratio is a good indicator for cell viability. High ATP and low ADP levels are usually found in proliferating cells. In contrast, decreased levels of ATP and increased levels of ADP are characteristic for apoptotic cells. In necrotic cells, the ADP levels are even higher, and ATP can be barely detected. The Bioluminescent Cell Viability Kit II utilizes bioluminescence to measure the ADP/ATP ratio providing an innovative system for fast, simultaneous screening of cell proliferation, apoptosis, necrosis, and growth arrest in mammalian cells. The ATP level is determined using the luciferin-luciferase reaction used in the Bioluminescent Cell Viability Kit I. The ADP level is then measured by its conversion to ATP, which is subsequently detected using the same luciferase-catalyzed reaction. The assay works with full automation for high throughput, detects as few as 100 mammalian cells per well, and offers highly consistent results with excellent correlation to other apoptosis markers (e.g., TUNEL-based or caspase assays).

Cell Proliferation Kit I (CFSE) PromoKine’s Cell Proliferation Kit I provides a convenient and sensitive way to determine cell proliferation or cell division. The assay is based on the amine-reactive dye CFDA SE (5-(and 6)-carboxyfluorescein diacetate, succinimidyl ester; also known as CFSE) which is non-fluorescent and can diffuse passively into cells. When it enters the cells, the acetate group is removed by intracellular esterases yielding the bright greenfluorescent dye carboxyfluorescein SE. Its succinimidyl ester (SE) reactive group reacts with intracellular amines and attaches the fluorescent dye to intracellular proteins, retaining it in the cells and allowing fixation with aldehyde fixatives. CFDA SE has been used as a tracer for several cell divisions as the label is stably inherited by daughter cells through successive cell divisions or after cell fusion and is not transferred to adjacent cells in a population. The kit is compatible with flow cytometry and microscopy and contains 10 single-use vials as well as a detailed protocol for cellular labeling.

https://www.promokine.info/shop Product

Size

Catalog Number

500 assays 2500 assays 10000 assays

PK-CA707-30025-0 PK-CA707-30025-1 PK-CA707-30025-2

1000 assays

PK-CA707-30026

Colorimetric Cell Viability Kit I (WST-8)

100 assays 500 assays

PK-CA705-CK04-100 PK-CA705-CK04

Colorimetric Cell Viability Kit II (WST-1)


PK-CA577-K304-500 PK-CA577-K304-2500

Colorimetric Cell Viability Kit III (XTT)


PK-CA20-300-200 PK-CA20-300-1000

Colorimetric Cell Viability Kit IV (MTT)


PK-CA707-30006-200 PK-CA707-30006



PK-CA577-K254-200 PK-CA577-K254-1000


200 assays

PK-CA577-K255

Cell Proliferation Kit I (CFSE)

10 x 50 µg

PK-CA707-30050

10 mg

PK-CA707-10054



Resazurin, sodium salt

Note: Please check our website regularly for new cell-based assays.

6 Cell Analysis

105

6.2 Live/Dead Cell Staining Kits

Monitoring of viable and dead cells Cell counting by fluorescence micros copy and fluor cytometry

Features

Simultaneous monitoring of viable and dead cells High fluorescent intensity Fast and convenient procedure Excitation and emission spectra are compatible with common excitation sources and filter sets

Live/Dead Cell Staining Kit II The Live/Dead Cell Staining Kit II utilizes two fluorescent dyes, Calcein-AM and

Calcein-AM R: acetoxymethyl

Ethidium homodimer III (EthD-III). Calcein-AM penetrates the cell membrane of living cells. Once inside, the cellular esterases present in living cells hydrolyze it to cell membrane-impermeable, green-fluorescent Calcein (emission maximum at 515 nm). In contrast, EthD-III can only pass through the disrupted membrane areas of dead cells where it intercalates with the DNA of the nucleus, emitting red light at 620 nm. Compared with PI, EthD-III is more stable and emits brighter fluorescent signals. Since both Calcein and EthD-III can be excited at 490 nm, a simultaneous monitoring of viable and dead cells is possible using a fluorescence microscope (see Fig. 4). At an excitation of 545 nm, the number of dead cells can be monitored.

Bacteria Live/Dead Staining Kit The Bacteria Live/Dead Staining Kit enables a two-color fluorescent staining on both live bacteria (green) and dead bacteria (red) using the two nucleic acid dyes DMAO and EthD-III. DMAO is a green-fluorescent dye which stains both live and dead bacteria with intact and damaged cell membranes while the red-fluorescing EthD-III only stains dead bacteria with damaged cell membranes. With an appropriate mixture of both dyes, viable bacteria with intact cell membranes are stained green, whereas bacteria with damaged cell membranes fluoresce red. The kit is applicable to most bacteria types for viability and cytotoxicity studies and is suitable for fluorescence microscopy and flow cytometry.

EthD-III

Esterase

Viable cell

DNA intercalation red fluorescence

Fig. 4: Double-staining of viable (green) and dead (red) cells using the PromoKine Live/Dead Cell Staining Kit II.

Calcein green fluorescence

Fig. 3: Assay mechanism of the PromoKine Live/Dead Cell Staining Kit II.


Size

Catalog Number

Live/Dead Cell Staining Kit II

300-1000 assays

PK-CA707-30002

Bacteria Live/Dead Staining Kit

100-1000 assays

PK-CA707-30027

Please see also the PromoKine Apoptotic/Necrotic/Healthy Cells Detection Kit on page 140 and our individual stains for viable and dead cells on page 110. Note: Please check our website regularly for new cell-based assays.

Cell Analysis

Applications

106

6 Cell Analysis

6.3 Cytotoxicity Kits Applications Cytotoxicity assays Chemosensitivity testing Toxicity screening

Cell Analysis

Features

Non-radioactive method Highly sensitive Easy and fast procedure Compatible with common plate readers

It oxidizes lactate to pyruvate, triggering the conversion of the yellow tetrazolium salt WST into the water-soluble red formazan dye. The increase in the amount of formazan produced in culture medium can be analyzed spectrophotometrically and correlates directly to the increase in the number of lysed cells. The kit can be used for many different in vitro cell systems to quantify cytotoxicity and cytolysis.

LDH Cytotoxicity Kit II

AK Cytotoxicity Kit

The colorimetric LDH Cytotoxicity Kit II provides a fast, sensitive, and precise alternative to the radioactive [3H]-thymidine and [51Cr] release assays. It is based on the activity of lactate dehydrogenase (LDH), a stable cytoplasmic enzyme that is present in all cells. Damaged cells rapidly release this enzyme into the surrounding medium. LDH activity is determined by a coupled enzymatic reaction.

The bioluminometric AK Cytotoxicity Kit is based on the measurement of adenylLDH

Lactate NAD+ Formazan (red)

ate kinase (AK), a ubiquitous protein present in all cells that is rapidly released into the culture medium upon damage to the plasma membrane. The simple one-step procedure involves two chemical reactions. The adenylate kinase release from the damaged cells catalyses the conversion of ADP to ATP. The luciferase catalyses a light-producing reaction between the ATP, oxygen, and luciferin. The light can be measured using a luminometer or beta counter. This highly sensitive assay allows the rapid quantitation of plasma membrane damage for evaluation of cell death or cytotoxicity events and it is well suited for automation and high throughput.

Pyruvate NADH + H+

Catalyst

WST-1 (yellow)

Fig. 5: Assay mechanism of the PromoKine LDH Cytotoxicity Kit II. https://www.promokine.info/shop Product

Size

Catalog Number


500 assays

PK-CA577-K313

AK Cytotoxicity Kit

500 assays

PK-CA577-K312

6.4 Senescence Detection Kit Applications Histochemical detection of senescent cells Cell senescence and oncology studies

Features High sensitivity and specificity Compatible with common lab equipment

Senescence is an arrested state in which the cells remain viable but are not stimulated to divide by serum or passage in culture. It is thought to be a tumor-suppressive mechanism and an underlying cause of aging. Senescent cells display an increase in size, senescence-associated expression of b-galactosidase (SA-b-Gal) activity, and altered patterns of gene expression. The PromoKine Senescence

Detection Kit has been designed to histochemically detect a specific senescence marker (SA-b-Gal activity) in cultured cells and tissue sections. The specific histochemical marker is only present in senescent cells and is not found in presenescent, quiescent, or immortal cells.

https://www.promokine.info/shop Product Senescence Detection Kit


Size

Catalog Number

250 assays

PK-CA577-K320

6 Cell Analysis

107

6.5 Cell-Based Reporter Assays

Monitoring and quantification of reporter gene expression (Luciferase, b-Galactosidase, GFP) Determination of transfection efficiencies

Features

Highly sensitive and reproducible Fast and convenient protocols Compatible with common plate readers

under these conditions, light emitted from the reaction is directly proportional to the number of luciferase enzyme molecules. The light emission from the luciferase reaction can be recorded using a luminometer. The Luciferase Reporter HTS Kits have been specifically designed for economic high-throughput applications using high-density microplates. In addition to the Firefly luciferase substrate D-Luciferin, we also provide a wide range of coelenterazine substrates for the Renilla luciferase catalysed reaction.

Luciferase Reporter Assay Kits GFP Reporter Assay Kit The PromoKine ready-to-use Luciferase Reporter Assays provide complete systems for rapid, convenient, and quantitative detection of luciferase activity in eukaryotic cells, e.g. in cells transfected with plasmids carrying the Firefly and/ or Renilla luciferase reporter genes. They are optimized to provide excellent sensitivity as well as consistent light output when working with multiple samples. Firefly luciferase catalyzes the oxidative carboxylation of luciferin, a reaction with the highest efficiency of any known bioluminescence reaction. Consequently, this sensitive reporter is ideally suited for studying gene regulation and function. Renilla luciferase is often used in conjunction with Firefly luciferase as a normalizing transfection control or for multiplexing transcriptional reporter assays. Both Firefly luciferase and Renilla luciferase show maximum light output when substrates and co-factors are present in large quantities. When assayed

GFP (Green Fluorescent Protein) is a fluorescent protein originally isolated from the jellyfish Aequorea victoria. GFP and its enhanced variant EGFP have been widely used as reporter proteins for in vivo monitoring of gene expression in a variety of cell types and organisms. Since GFP requires no additional substrates or cofactors, GFP fluorescence can easily be detected using a fluorescence microscope. However, most imaging studies of GFP are qualitative. Quantitative analyses of GFP expression levels in cells or tissues are more informative and have wider applications. The PromoKine GFP Reporter Assay Kit was designed for use in a 96-well microplate format (Ex/Em = 488 nm/507nm). Cells or tissues can be homogenized directly in the provided GFP assay buffer and the supernatant of the lysed cells/ tissues is transfered to the microwell plates. The amount of GFP is determined

by comparing its fluorescence with that of a GFP standard. The kit can detect a wide range of GFP concentrations (0.0110 μg/ml). Furthermore, a GFP quench solution is provided for determining autofluorescence levels of cell or tissue extracts.

Beta-Galactosidase Kits The b-Galactosidase Staining Kit and the b-Galactosidase Reporter Assay Kit were designed to detect and quantify b-Gal activity in cells transfected with a plasmid carrying the frequently-used LacZ reporter gene. This gene encodes b-galactosidase, an enzyme that is very stable and resistant to proteolytic degradation. When expressed in transfected cells, it converts a colorless substrate (e.g., CPRG, X-Gal, or ONPG) into a colored reaction product that can be easily detected microscopically (b-Galactosidase Staining Kit) or quantified using a spectrophotometer (b-Galactosidase Reporter Assay Kit). In addition, PromoKine provides a range of chromogenic beta-Galactosidase substrates.

Fig. 6: b-Gal staining of CHO-K1 cells transfected with the pPK-CMV-R1 reporter vector expressing the lacZ gene (see page 166).


Size

Catalog Number

Luciferase Reporter Assay Kit I (Firefly)


PK-CA707-30003-1 PK-CA707-30003-2

Luciferase Reporter Assay Kit II (Renilla)


PK-CA707-30004-1 PK-CA707-30004-2

Luciferase Reporter Assay Kit III (Firefly & Renilla)


PK-CA707-30005-1 PK-CA707-30005-2

Luciferase Reporter HTS Kit (Firefly)

4 ml 10 ml 100 ml 10 x 100 ml

PK-CA707-30028-0 PK-CA707-30028-1 PK-CA707-30028-2 PK-CA707-30028-3

Cell Analysis

Applications

108

6 Cell Analysis


Size

Catalog Number

beta-Galactosidase Reporter Assay Kit

500 assays

PK-CA708-10100

beta-Galactosidase Staining Kit

200 assays

PK-CA577-K802

GFP Reporter Assay Kit

100 assays

PK-CA577-K815

D-Luciferin, Free Acid

10 mg 50 mg 1g

PK-CA707-10100 PK-CA707-10100-1 PK-CA707-10100-2

D-Luciferin, Potassium Salt

10 mg 50 mg 1g

PK-CA707-10101 PK-CA707-10101-1 PK-CA707-10101-2

D-Luciferin, Sodium Salt

10 mg 50 mg 1g

PK-CA707-10102 PK-CA707-10102-1 PK-CA707-10102-2

5 mg

PK-CA707-10103

Coelenterazine (native)

50 μg

PK-CA707-10110

Coelenterazine h

50 μg

PK-CA707-10111

Coelenterazine cp

50 μg

PK-CA707-10112

Coelenterazine hcp

50 μg

PK-CA707-10113

Coelenterazine f

50 μg

PK-CA707-10114

Coelenterazine n

50 μg

PK-CA707-10115

Coelenterazine ip

50 μg

PK-CA707-10116

Coelenterazine fcp

50 μg

PK-CA707-10117

Coelenterazine i

50 μg

PK-CA707-10121

Coelenterazine, 2-methyl

50 μg

PK-CA707-10122

Coelenterazine 400a (also known as DeepBlue C, a trademark of Perkin Elmer)

50 μg

PK-CA707-10125

1 Kit

PK-CA707-10123

X-Gal

100 mg 1g 5g

PK-CA707-10011 PK-CA707-10011-1 PK-CA707-10011-2

Red-beta-D-Gal

100 mg

PK-CA707-10012

Rose-beta-D-Gal

100 mg

PK-CA707-10013

25 mg

PK-CA707-10014

Cell Analysis

Firefly Luciferase Substrates

DMNPE-caged D-Luciferin Renilla Luciferase Substrates

Coelenterazine Sampler Kit Chromogenic beta-Galactosidase Substrates

Purple-beta-D-Gal Green-beta-D-Gal Chromogenic beta-Galactosidase Substrate Sampler Kit

25 mg

PK-CA707-10015

1 kit (5 x 5 mg)

PK-CA707-10024

Please see page 166 and www.promokine.info for luciferase, beta-Gal, and GFP reporter vectors. Note: Please check our website regularly for new cell-based assays.

6 Cell Analysis

109

6.6 Cell Staining Reagents and Related Products

Detection of cellular structures Fluorescence microscopy, Flow cytometry

Features High quality Thoroughly tested Highly sensitive Fluorescent dyes offer extraordinary sensitivity and extended possibilities for the visualization and analysis of intracellular processes and cell structures such as the cell membrane, mitochondria, the nucleus, and the cytoskeleton. Some dyes

are also suited for staining only viable or dead cells. That is an important feature for elucidating the effects of certain agents on cell viability. PromoKine provides a select choice of reagents for staining different cell organelles and structures as well as for determining processes like membrane potential changes. Some of these reagents are available as esters, which pass through the membranes of viable cells. Once inside, the ester group is hydrolyzed by cellular esterases, and the dye can accumulate in the cell. Some dyes are nonfluorescent in their ester form but fluoresce strongly when the ester group has been removed.

Fig. 7: Primary human fibroblasts (NHDF, see page 27) stained with Calcein and counterstained with DAPI.


Product Description

Size

Catalog Number

1 mg

PK-CA707-40037

10 ml (10 mg/ml)

PK-CA707-40039

10 mg

PK-CA707-40011

10 mg in 1 ml

PK-CA707-40043

10 mg

PK-CA707-40009

Nucleus Stains 7-AAD (7-aminoactinomycin D)

Fluorescent DNA intercalator

Acridine Orange (AO) Solution

Nucleus stain (DNA/RNA)

DAPI

Nucleus stain (DNA)

DAPI Solution

Nucleus stain (DNA); aqueous solution

DAPI, dilactate

4’,6-diamidino-2-phenylindole, dilactate

DMAO

Nucleus stain (DNA); 2 mM solution in DMSO

EB (ethidium bromide) Solution

Dead cell stain (DNA), aqueous solution

Ethidium Homodimer I

Nucleus stain (DNA); also known as EthD-1, or EtDi

Ethidium Homodimer III

Nucleus stain (DNA)

Ethidium Homodimer III, 1 mM in DMSO

Nucleus stain (DNA); in DMSO

Hoechst 33258

Nucleus stain (DNA)

Hoechst 33258 Solution


Hoechst 33342

Nucleus stain (DNA)



Propidium Iodide (PI)

Dead cell stain (DNA)

Propidium Iodide (PI) Solution

Dead cell stain (DNA); aqueous solution

1 ml

PK-CA707-40012

10 ml (10 mg/ml)

PK-CA707-40042

1 mg

PK-CA707-40010

1 mg

PK-CA707-40050

200 µl

PK-CA707-40051

100 mg

PK-CA707-40045

10 ml (10 mg/ml)

PK-CA707-40044

100 mg

PK-CA707-40047

10 ml (10 mg/ml)

PK-CA707-40046

100 mg

PK-CA707-40016

10 ml (1 mg/ml)

PK-CA707-40017

Mitochondria Stains DASPEI

2-(4-(dimethylamino)styryl)-N-ethylpyridinium iodide; for staining of mitochondria in live cells

100 mg

PK-CA707-70018

Dihydrorhodamine 123

Reduced form of Rhodamine 123; mitochondria stain

10 mg

PK-CA707-10055

Dihydrorhodamine 123, dihydrochloride salt

Reduced form of Rhodamine 123; mitochondria stain

10 mg

PK-CA707-10056

DiOC6(3)

3,3’-dihexyloxacarbocyanine, iodide; membrane potential sensitive dye

100 mg

PK-CA707-70009

Cell Analysis

Applications

110

6 Cell Analysis

Cell Analysis


Product Description

Size

Catalog Number

JC-1 chloride

Stains mitochondria in living cells in a membrane potential-dependent function

5 mg

PK-CA707-70011

JC-1 iodide

Stains mitochondria in living cells in a membrane potential-dependent function

5 mg

PK-CA707-70014

Nonyl Acridine Orange (or NAO)

Mitochondria stain; not dependent on mitochondrial membrane potential

50 mg

PK-CA707-70012

MitoGreen

Green-fluorescent mitochondrial dye

20 x 50 µg

PK-CA707-70054

MitoRed

Red-fluorescent mitochondrial dye

20 x 50 µg

PK-CA707-70055

Rhodamine 123

Green-fluorescent mitochondria stain

50 mg

PK-CA707-70010

Tetrabromorhodamine 123, bromide

Green-fluorescent mitochondria stain

5 mg

PK-CA707-70013

TMRE

Tetramethylrhodamine ethyl ester, perchlorate; membrane potential sensitive dye

25 mg

PK-CA707-70016

TMRM

Tetramethylrhodamine methyl ester, perchlorate; membrane potential sensitive dye

25 mg

PK-CA707-70017

100 mg

PK-CA707-70009

ER Stains DiOC6(3)

3,3’-dihexyloxacarbocyanine, iodide; membrane potential sensitive dye

Viable Cell Stains BCECF

Viable cell stain (cytosol)

1 mg

PK-CA707-51010

BCECF AM

Viable cell stain (cytosol); acetoxymethyl ester

10 x 100 µg 20 x 50 µg 1 mg

PK-CA707-51011 PK-CA707-51011-1 PK-CA707-51012

BCECF AM Solution

Viable cell stain (cytosol); acetoxymethyl ester; DMSO solution

1 mg in 1 ml

PK-CA707-51009

Calcein AM


20 x 50 µg 1 mg

PK-CA707-80011-3 PK-CA707-80011

Calcein AM Solution, 1 mg/ml in anhydrous DMSO

Viable cell stain (cytosol); acetoxymethyl ester, DMSO solution

1 mg in 1 ml

PK-CA707-80011-2

Calcein AM Solution, 4 mM DMSO solution

Viable cell stain (cytosol); acetoxymethyl ester, 4 mM DMSO solution

100 µl

PK-CA707-80011-1

Calcein, high purity

Viable cell stain (cytosol); ultra-pure

100 mg

PK-CA707-80013

CFSE; 5(6)-CFDA, SE

5-(and-6)-carboxyfluorescein diacetate, succinimidyl ester; viable cell stain (cytosol)

25 mg

PK-CA707-90041

Hoechst 33258

Nucleus stain (DNA)

100 mg

PK-CA707-40045



10 ml (10 mg/ml)

PK-CA707-40044

Hoechst 33342

Nucleus stain (DNA)

100 mg

PK-CA707-40047



10 ml (10 mg/ml)

PK-CA707-40046



10 ml (10 mg/ml)

PK-CA707-40042


Nucleus stain (DNA); also known as EthD-1, or EtDi

1 mg

PK-CA707-40010


Nucleus stain (DNA)

1 mg

PK-CA707-40050

Ethidium Homodimer III, 1 mM in DMSO

Nucleus stain (DNA); in DMSO

200 µl

PK-CA707-40051


Dead cell stain (DNA)



Trypan Blue Solution (0.5%)

Dead cell stain; tetrasodium salt, 0.5% solution

Dead Cell Stains

100 mg

PK-CA707-40016

10 ml (1 mg/ml)

PK-CA707-40017

10 ml

PK-CA902-1209

6 Cell Analysis

111


Product Description

Size

Catalog Number

Blue-fluorescent Cytoplasmic Membrane Staining Kit

Convenient and sensitive cell staining with low cytotoxicity

50 assays

PK-CA707-30024

Green-fluorescent Cytoplasmic Membrane Staining Kit


1 ml

PK-CA707-30021

Orange-fluorescent Cytoplasmic Membrane Staining Kit


1 ml

PK-CA707-30022

Red-fluorescent Cytoplasmic Membrane Staining Kit


1 ml

PK-CA707-30023

Lipophilic Carbocyanine Dyes DiD (DiIC18[5])

1,1'-dioctadecyl-3,3,3',3'- tetramethylindodicarbocyanine, 4-chlorobenzenesulfonate salt

50 mg

PK-CA707-60014

DiI (or DiIC18[3])

1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate

50 mg

PK-CA707-60010

DiI in vegetable oil

Optimized DiI formulation for microinjection

0.5 ml

PK-CA707-60018

Dilinoleyl DiI

1,1'-dilinoleyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (also called Fast DiI, a trademark of Molecular Probes)

5 mg

PK-CA707-60034

Dilinoleyl DiO

1,1'-dilinoleyl-3,3'-oxacarbocyanine perchlorate (also called Fast DiO, a trademark of Molecular Probes)

5 mg

PK-CA707-60035

DiO (or DiOC18[3])

3,3'-dioctadecyloxacarbocyanine, perchlorate

50 mg

PK-CA707-60011

DiOC14(3), hydroxyethanesulfonate

3,3'-ditetradecyloxacarbocyanine, hydroxyethanesulfonate

50 mg

PK-CA707-60012

DiOC16(3)

Lipophilic cyanine dye for staining cytoplasmic membranes

25 µg

PK-CA707-60038

DiR (DiIC18[7])

1,1'-dioctadecyl-3,3,3',3'-tetramethyl indotricarbocyanine Iodide

25 mg

PK-CA707-60017

Neuro-DiI

Improved alternative to DiI

25 mg

PK-CA707-60016

Neuro-DiI in vegetable oil

Optimized formulation for microinjection

0.2 ml

PK-CA707-60020

Neuro-DiO

Improved alternative to DiO

25 mg

PK-CA707-60015

Neuro-DiO in vegetable oil

Optimized formulation for microinjection

0.2 ml

PK-CA707-60019

Biotin-DHPE

N-(biotinoyl)-1,2-dihexadecanoyl-sn-glycero-3phosphoethanolamine, triethylammonium salt

10 mg

PK-CA707-60022

Biotin-X-DHPE

N-((6-(biotinoyl)amino)hexanoyl)-1,2-dihexadecanoylsn-glycero-3-phosphoethanolamine, triethylammonium salt

5 mg

PK-CA707-60023

Fluorescein-DHPE

N-(fluorescein-5-thiocarbamoyl)-1,2-dihexadecanoyl-snglycero-3-phosphoethanolamine, triethylammonium salt

5 mg

PK-CA707-60024

NBD-DHPE

N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine, triethylammonium salt

10 mg

PK-CA707-60025

Rhodamine-DHPE

N-(lissamine rhodamine B sulfonyl)-1,2-dihexadecanoylsn-glycero-3-phosphoethanolamine, triethylammonium salt

5 mg

PK-CA707-60026

Texas Red-DHPE

N-(Texas Red sulfonyl)-1,2-dihexadecanoyl-sn-glycero3-phosphoethanolamine, triethylammonium salt

1 mg

PK-CA707-60027

TRITC-DHPE

N-(teramethylrhodamine-6-thiocarbamoyl)-1,2dihexadecanoyl-sn-glycero-3-phosphoethanolamine, triethylammonium salt

1 mg

PK-CA707-60028

Lipophilic styryl dye with an amine group at the hydrophilic head

1 mg

PK-CA707-70051

Phospholipid Membrane Probes

Other Membrane Dyes AM3-25

Cell Analysis

Cytoplasmic Membrane Staining

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6 Cell Analysis

Cell Analysis


Product Description

Size

Catalog Number

Biotin-AM3-25

AM3-25 conjugate with biotin that can be coupled to avidin or streptavidin

1 mg

PK-CA707-60032

DiA

4-(4-dihexadecylaminostyryl)-N-methylpyridinium iodide

50 mg

PK-CA707-60013

DiB

Lipophilic blue fluorescent dye useful for staining cytoplasmic membranes

10 µg

PK-CA707-60036

DPA

Nonfluorescent anionic membrane dye that can be used as a FRET quencher for a variety of fluorescent donor dyes

25 µg

PK-CA707-60037

NBD C6-ceramide

6-((N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino) hexanoyl)sphingosine

1 mg

PK-CA707-60030

NBD C6-sphingomyelin

6-((N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino) hexanoyl)sphingosyl phosphocholine

1 mg

PK-CA707-60031

Octadecyl Rhodamine B Chloride (R18)

Useful for membrane fusion assays

10 mg

PK-CA707-60033

Nerve Terminal Staining Kits Nerve Terminal Green Staining Kit I

Kit contains 5 x 1 mg SynapseGreen C4 and 250 mg ADVASEP-7

1 set

PK-CA707-70030

Nerve Terminal Green Staining Kit II

Kit contains 1 mg AM1-43 and 100 mg ADVASEP-7

1 set

PK-CA707-70031

Nerve Terminal Green Staining Kit III

Kit contains 1 mg AM1-43 and 100 mg SCAS

1 set

PK-CA707-70031-1

Nerve Terminal Green Staining Kit IV

Kit contains 5 x 1 mg SynapseGreen C4 and 100 mg Sulforhodamine 101

1 set

PK-CA707-70032

Nerve Terminal Red Staining Kit I

Kit contains 5 x 1 mg SynapseRed C2 and 250 mg ADVASEP-7

1 set

PK-CA707-70034

Nerve Terminal Red Staining Kit II

Kit contains 5 x 1 mg SynapseRed C2 and 100 mg Sulforhodamine 101

1 set

PK-CA707-70035

250 mg

PK-CA707-70029

Nerve Terminal Probes & Related Reagents ADVASEP-7 (sulfobutyl b-cyclodextrin)

For background fluorescence reduction

Alpha-Bungarotoxin (from Bungarus multicinctus)

Acetylcholine receptor inhibitor

1 mg

PK-CA707-00010-1

AM1-43

Fixable, activity-dependent fluorescent nerve terminal probe

1 mg

PK-CA707-70024

AM1-44

Nerve Terminal Probe

1 mg

PK-CA707-70038

AM2-10


1 mg

PK-CA707-70036

AM3-25


1 mg

PK-CA707-70051

AM4-64


1 mg

PK-CA707-70025

AM4-66


1 mg

PK-CA707-70050

Biotin-XX-a-Bungarotoxin

Can be used to isolate and/or image the distribution of the nicotinic acetylcholine receptor at the neuromuscular junctions

500 µg

PK-CA707-00017

Fluorescein-a-Bungarotoxin

Can be used to image the distribution of the nicotinic acetylcholine receptor at the neuromuscular junctions

500 µg 10 x 50 µg

PK-CA707-00011 PK-CA707-00013

SCAS

4-sulfonato calix[8]arene, sodium salt; for background fluorescence reduction

100 mg

PK-CA707-70037

Sulforhodamine 101-aBungarotoxin

Also known as Texas Red-a-Bungarotoxin

500 µg 10 x 50 µg

PK-CA707-00015 PK-CA707-00016

SynapseGreen C1


5 mg 5 x 1 mg

PK-CA707-70042 PK-CA707-70043

SynapseGreen C18 (FM3-25)

Nerve Terminal Probe (also known as FM3-25, a trademark of Molecular Probes, Inc.)

5 mg 5 x 1 mg

PK-CA707-70048 PK-CA707-70049



5 mg 5 x 1 mg

PK-CA707-70044 PK-CA707-70045

6 Cell Analysis

113

Product

Product Description

Size

Catalog Number

SynapseGreen C3


5 mg 5 x 1 mg

PK-CA707-70023 PK-CA707-70026



5 mg 5 x 1 mg

PK-CA707-70020 PK-CA707-70022



5 mg 5 x 1 mg

PK-CA707-70046 PK-CA707-70047

SynapseRed C1


5 mg 5 x 1 mg

PK-CA707-70040 PK-CA707-70041

SynapseRed C2 (FM4-64)


5 mg 5 x 1 mg

PK-CA707-70021 PK-CA707-70027

SynapseRed C2M (FM5-95)


5 mg 5 x 1 mg

PK-CA707-70028 PK-CA707-70019

Tetramethylrhodaminea-Bungarotoxin

Can be used to image the distribution of the nicotinic acetylcholine receptor at the neuromuscular junctions

500 µg 10 x 50 µg

PK-CA707-00012 PK-CA707-00014

Membrane Potential Sensitive Dyes DiO/DPA Membrane Potential Detection Kit

For fluorescent detection of cytoplasmic membrane potential changes

1 kit

PK-CA707-30037

Di-12-ANEPPQ

Fast-responding membrane potential dye

5 mg

PK-CA707-61015

Di-2-ANEPEQ (JPW1114)

Fast-responding membrane potential dye; also called JPW1114

5 mg

PK-CA707-61013

Di-4-ANEPPS


5 mg

PK-CA707-61010

Di-8-ANEPPQ


5 mg

PK-CA707-61014

Di-8-ANEPPS


DiBAC4(3)

Bis-(1,3-dibarbituric acid)-trimethine oxanol; membrane potential sensitive dye

DiIC1(5)

5 mg

PK-CA707-61012

25 mg

PK-CA707-61011

1,1'-dimethyl-3,3,3',3'-tetramethylindodicarbocyanine, iodide

100 mg

PK-CA707-70015

DiOC2(3)

3,3’-diethyloxacarbocyanine, iodide; membrane potential sensitive dye

100 mg

PK-CA707-70008

DiOC5(3)

3,3’-dipentyloxacarbocyanine, iodide; membrane potential sensitive dye

100 mg

PK-CA707-70007

DiOC6(3)

3,3’-dihexyloxacarbocyanine, iodid; membrane potential sensitive dye

100 mg

PK-CA707-70009

RH237

Fast-responding potentiometric probe

5 mg

PK-CA707-61018

RH414


5 mg

PK-CA707-61016

RH421


25 mg

PK-CA707-61017

RH795


5 mg

PK-CA707-61019

TMRE

Tetramethylrhodamine ethyl ester, perchlorate; membrane potential sensitive dye

25 mg

PK-CA707-70016

TMRM

Tetramethylrhodamine methyl ester, perchlorate; membrane potential sensitive dye

25 mg

PK-CA707-70017

Cytosolic Tracers and Related Reagents Biocytin (e-biotinoyl–L-lysine)

Can be introduced into cells by microinjection

100 mg

PK-CA707-90055

Biocytin-Hydrazide

Can be introduced into cells by microinjection; aldehyde-fixable

25 mg

PK-CA707-90060

Biotin-4-Fluorescein

Can be used as polar tracer to study cell morphology

10 mg

PK-CA707-90062

Biotin-Ethylenediamine, hydrobromide

Equivalent to Neurobiotin; transneuronal tracer

25 mg

PK-CA707-90057

Biotin-Ethylenediamine, hydrochloride

Functionally equivalent to Neurobiotin; transneuronal tracer

25 mg

PK-CA707-90075

Biotin-Rhodamine 110

Can be used as polar tracer to study cell morphology

5 mg

PK-CA707-80022

Cell Analysis

https://www.promokine.info/shop

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6 Cell Analysis

Cell Analysis


Product Description

Size

Catalog Number

Calcein AM


1 mg 20 x 50 µg

PK-CA707-80011 PK-CA707-80011-3

Calcein AM Solution, 1 mg/ml in anhydrous DMSO


1 ml

PK-CA707-80011-2

Calcein AM Solution, 4 mM DMSO solution


100 µl

PK-CA707-80011-1


Viable cell stain (cytosol); ultra-pure

100 mg

PK-CA707-80013

DPA/Terbium for membrane fusion assay

Can be used for vesicle fusion assays

1 set

PK-CA707-80104

DPX

Can be used to study membrane fusion or permeability

500 mg

PK-CA707-80012

Fluorescein-Biotin

5-((N-(5-(N-(6-(biotinoyl)amino)hexanoyl)amino)pentyl) thioureidyl) fluorescein

5 mg

PK-CA707-80019

Hydroxystilbamidine

Also known as FluoroGoldTM

10 mg

PK-CA707-80014

Hydroxystilbamidine

TM

4% in H2O; also known as FluoroGold

200 µl

PK-CA707-80023

Lucifer Yellow Cadaverine

Can be used as fixable fluorescent tracer

25 mg

PK-CA707-80018

Lucifer Yellow Cadaverine Biotin-X, dipotassium salt

Useful tracer for studying neuronal morphology

10 mg

PK-CA707-80017

Lucifer Yellow CH lithium salt

Useful polar tracer for studying neuronal morphology

25 mg

PK-CA707-80015

Lucifer Yellow CH potassium salt

Useful polar tracer for studying neuronal morphology

25 mg

PK-CA707-80016

RuBP-2S

Ruthenium(bis(bathophenanthroline))(bathophenanthrolinedisulfonate

5 mg

PK-CA707-80024

RuBP-4S

Ruthenium(bathophenanthroline)(bis(bathophenanthroli nedisulfonate)), disodium salt

5 mg

PK-CA707-80025

RuBPS

Ruthenium(tris(bathophenanthrolinedisulfonate), tetrasodium salt

5 mg

PK-CA707-80021

SDIP/Europium for membrane fusion assay

Can be used for vesicle fusion assays

1 set

PK-CA707-80105

Sulforhodamine 101

Useful polar tracer for studying neuronal morphology and cell-cell communication

500 mg

PK-CA707-80101

Sulforhodamine B


5g

PK-CA707-80100

Sulforhodamine G


5g

PK-CA707-80102

10 ml

PK-CA707-90082

Related Products DMSO, anhydrous

Dimethylsulfoxide, anhydrous

Pluronic F-127

Nonionic detergent useful for solubilizing large dye molecules such as Calcein/AM, Fura-2/AM, Fluo-3/AM, Indo-1/AM and Rhod-2/AM to facilitate cell loading

1g

PK-CA707-59000

Pluronic F-127, 20% solution in DMSO


1 ml

PK-CA707-59004

Pluronic F-127, 10% in H2O


30 ml

PK-CA707-59005

Note: Please check our website regularly for new cell stains.

6 Cell Analysis

115

6.7 Intracellular Indicators and Related Products

6.7.1 Calcium Indicators Applications

Intracellular calcium signaling studies Investigation of signal transduction and apoptotic cascades Neuroscience research

Features High quality and sensitivity

Derivatives for special applications available Compatible with standard fluores cence equipment Calcium ions are important second messengers in cells. They are involved in muscle contraction, nerve signal transmission, immune cell activation, hormone secretion, etc. Fluorescent calcium indicators selectively bind to calcium ions and show a spectral response upon binding Ca2+, i.e., fluorescence wavelength or intensity are altered. This shift of fluorescence wavelength and change of intensity upon calcium chelation can be monitored using fluorescence microscopy. PromoKine offers a wide range of fluorescent calcium indicators for detecting changes in the intracellular Ca2+ concentration, including Fura-2, Fluo-3, Rhod2, and Indo-1 (as cell-impermeable salts or cell-permeable AM esters) as well as their low-affinity derivatives Fura-2FF,

n Calcium Indicators Product

Fluo-3FF, and Indo-1FF which are particularly useful for certain applications (e.g., spatial and functional characterization of Ca2+ stores or detection of NMDA-induced neuronal Ca2+ fluxes). PromoKine also provides a range of nonfluorescent calcium chelators as well as coelenterazine derivatives which are effective luminescent calcium indicators.

6.7.2 Other Indicators Applications pH determination Iron, zinc, and chloride ion detection

Features High quality and sensitivity In addition to our calcium probes, PromoKine also offers pH indicators as well as indicators for iron, zinc, and chloride ion detection.

https://www.promokine.info/shop Product Description

Size

Catalog Number

Fluorescent Calcium Indicators Fluo-3 (pentaammonium salt)

Fluorescent calcium indicator; Ex/Em: 490-506/526

1 mg

PK-CA707-50010

Fluo-3 (pentapotassium salt)


1 mg

PK-CA707-50011

Fluo-3 (pentasodium salt)


1 mg

PK-CA707-50012

Fluo-3 AM

Fluorescent calcium indicator (acetoxymethyl ester)

10 x 100 µg 20 x 50 µg 1 mg

PK-CA707-50013 PK-CA707-50016 PK-CA707-50014

Fluo-3 AM Solution

Fluorescent calcium indicator (acetoxymethyl ester; 1 mM in anhydrous DMSO)

1 ml

PK-CA707-50015

Fura-2 (pentaammonium salt)


1 mg

PK-CA707-50030

Fura-2 (pentapotassium salt)


1 mg

PK-CA707-50031

Fura-2 (pentasodium salt)


1 mg

PK-CA707-50032

Fura-2 AM


10 x 100 µg 20 x 50 µg 1 mg

PK-CA707-50033 PK-CA707-50033-1 PK-CA707-50034

Fura-2 AM Solution

Fluorescent calcium indicator (acetoxymethyl ester; 1 mM in anhydrous DMSO)

1.0 ml

PK-CA707-50029

Indo-1 (pentaammonium salt)

Fluorescent calcium indicator; Ex/Em: 330/405-485

1 mg

PK-CA707-50040

Indo-1 (pentapotassium salt)


1 mg

PK-CA707-50041

Indo-1 (pentasodium salt)


1 mg

PK-CA707-50042

Indo-1 AM


10 x 100 µg 20 x 50 µg 1 mg

PK-CA707-50043 PK-CA707-50043-1 PK-CA707-50044

Rhod-2 (triammonium salt)

Fluorescent calcium indicator; Ex/Em: 533-556/~578

1 mg

PK-CA707-50020

Cell Analysis

Signal transduction often starts with the activation of a receptor on the cell surface which subsequently triggers a cascade of second messengers in the cell. The change of intracellular ion concentration is a crucial step that helps second messengers transmit information. Consequently, monitoring and control of intracellular ion concentration is a prerequisite understanding second messengers and signal transduction in living cells.

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6 Cell Analysis


Product Description

Size

Catalog Number

Rhod-2 (tripotassium salt) Rhod-2 (trisodium salt)


1 mg

PK-CA707-50021


1 mg

PK-CA707-50022

Rhod-2 AM


10 x 100 µg 1 mg

PK-CA707-50023 PK-CA707-50024

Cell Analysis

Fluorescent High Calcium Indicators 5,5'-dibromo-Bapta (tetrapotassium salt)

Fluorescent calcium indicator

100 mg

PK-CA707-50004

5,5'-difluoro-Bapta (tetrapotassium salt)


100 mg

PK-CA707-50006

5,5'-difluoro-Bapta AM


25 mg

PK-CA707-50005

1 mg

PK-CA707-50045

500 μg

PK-CA578-F1046

Bis-Fura-2, hexapotassium salt


Fura-2FF (K+ Salt)

Difluorinated derivative of Fura-2 (K+ Salt)

Fura-2FF AM

Difluorinated derivative of Fura-3 (acetoxymethyl ester)

500 μg

PK-CA578-F1044

Furaptra, AM ester

Fluorescent calcium indicator (also known as Mag-Fura-2)

10 x 100 µg 20 x 50 µg 1 mg

PK-CA707-50037 PK-CA707-50039 PK-CA707-50038

Furaptra, tetrapotassium salt


1 mg

PK-CA707-50035

Furaptra, tetrasodium salt


1 mg

PK-CA707-50036

Indo-1FF (K Salt)

Difluorinated derivative of Indo-1 (K Salt)

500 μg

PK-CA578-I1086

Indo-1FF AM

Difluorinated derivative of Indo-1 (acetoxymethyl ester)

500 μg

PK-CA578-I1084

+

+

Fluorescent Near-Membrane Calcium Indicators FFP-18 AM

Fluorescent near-membrane calcium indicator (Fura-derivative; acetoxymethyl ester)

1 mg

PK-CA578-F1052-1

FIP-18 AM

Fluorescent near-membrane calcium indicator (Indo-derivative; acetoxymethyl ester)

500 ug

PK-CA578-F1054

5,5'-dibromo-Bapta (tetrapotassium salt)


100 mg

PK-CA707-50004

5,5'-dimethyl BAPTA, AM ester


25 mg

PK-CA707-50007

5,5'-dimethyl BAPTA, tetrapotassium salt


100 mg

PK-CA707-50008

Bapta (tetracesium salt)

Non-fluorescent calcium chelator

1g

PK-CA707-50001

Bapta (tetrapotassium salt)


1g

PK-CA707-50002

Bapta (tetrasodium salt)


1g

PK-CA707-50003

Bapta AM

Non-fluorescent calcium chelator (acetoxymethyl ester)

25 mg 20 x 1 mg

PK-CA707-50000 PK-CA707-50000-1

Bapta-FF (free acid)

Difluorinated derivative of BAPTA

10 mg

PK-CA578-B1091

Bapta-FF AM

Difluorinated derivative of BAPTA (acetoxymethyl ester)

10 mg

PK-CA578-B1093


Coelenterazine; luminescent calcium indicator & luciferase substrate

50 μg

PK-CA707-10110

Coelenterazine 400a

Coelenterazine; luminescent calcium indicator & luciferase substrate (also known as DeepBlue C, a trademark of Perkin Elmer)

50 μg

PK-CA707-10125

Coelenterazine cp

Coelenterazine derivative; luminescent calcium indicator & luciferase substrate

50 μg

PK-CA707-10112

Coelenterazine f


50 μg

PK-CA707-10114

Coelenterazine fcp


50 μg

PK-CA707-10117

Calcium Chelators

Luminescent Calcium Indicators

6 Cell Analysis

117

Product

Product Description

Size

Catalog Number

Coelenterazine h


50 μg

PK-CA707-10111

Coelenterazine hcp


50 μg

PK-CA707-10113

Coelenterazine i


50 μg

PK-CA707-10121

Coelenterazine ip


50 μg

PK-CA707-10116

Coelenterazine n


50 μg

PK-CA707-10115



50 μg

PK-CA707-10122

Coelenterazine Sampler Kit

Coelenterazine and derivatives thereof

1 Kit

PK-CA707-10123

n Other Indicators Product

https://www.promokine.info/shop Product Description

Size

Catalog Number

Zn Indicators Zinquin, ethyl ester

Blue-fluorescent Zn indicator

5 mg

PK-CA707-52020

Zinquin, free acid

Blue-fluorescent Zn indicator

5 mg

PK-CA707-52022

100 mg

PK-CA707-52011

Cl Indicators MQAE

N-(ethoxycarbonylmethyl)-6-methoxyquinolinium bromide; fluorescent chloride ion indicator

SPQ

6-methoxy-N-(3-sulfopropyl)quinolinium; fluorescent chloride ion indicator

50 mg

PK-CA707-52010

Tetrakis-(2-pyridylmethyl)ethylenediamine; heavy metal chelator

100 mg

PK-CA707-59003

5(6)-CFDA

5-(and-6)-carboxyfluorescein diacetate; fluorescent pH indicator

100 mg

PK-CA707-51014

5(6)-CFDA, SE (CFSE)

5-(and-6)-carboxyfluorescein diacetate, succinimidyl ester

25 mg

PK-CA707-90041

5-(and-6)-Carboxy-2',7'dichlorofluorescein

Fluorescent pH indicator

200 mg

PK-CA707-51015

5-(and-6)-Carboxy-2',7'dichlorofluorescein diacetate

Fluorescent pH indicator; cell membrane-permeant

100 mg

PK-CA707-51016

5-(and-6)-Carboxy-2',7'dichlorofluorescein diacetate, succinimidyl ester (CDFDA, SE)

Fluorescent pH indicator; cell membrane-permeant

25 mg

PK-CA707-90040

5-(and-6)-Carboxyfluorescein


100 mg

PK-CA707-51013

5-Carboxy-2',7'dichlorofluorescein

Fluorescent pH indicator (single isomer)

100 mg

PK-CA707-51017

5-Carboxy-2',7'dichlorosulfonefluorescein

Fluorescent pH indicator (single isomer)

10 mg

PK-CA707-51023

5-CFDA

5-Carboxyfluorescein diacetate (single isomer); fluorescent pH indicator

100 mg

PK-CA707-51018

5-FAM

5-Carboxyfluorescein; fluorescent pH indicator

100 mg

PK-CA707-51019

6-CFDA

6-Carboxyfluorescein diacetate (single isomer); fluorescent pH indicator

100 mg

PK-CA707-51021

Heavy Metal Ion Indicators TPEN pH Indicators

Cell Analysis


118

6 Cell Analysis

Cell Analysis


Product Description

Size

Catalog Number

6-FAM

6-carboxyfluorescein; fluorescent pH indicator

BCECF

2',7'-bis-(carboxyethyl)-5-(and-6)-carboxyfluorescein; fluorescent pH indicator

100 mg

PK-CA707-51020

1 mg

PK-CA707-51010

BCECF AM

Fluorescent pH indicator; acetoxymethyl ester

10 x 100 µg 20 x 50 µg 1 mg

PK-CA707-51011 PK-CA707-51011-1 PK-CA707-51012

BCECF AM Solution

Fluorescent pH indicator; acetoxymethyl ester (1 mg/ml in dry DMSO)

1 ml

PK-CA707-51009

Flubida-2


5 mg

PK-CA707-51022

n Related Products


Product

Product Description

4-Bromo A-23187 free acid

For in situ calibration of fluorescent calcium indicators

A-23187 free acid

A-23187 Free Acid (calcimycin; or Calcium Ionophore III)

Calcium Calibration Buffer Kit

For calibration of fluorescent calcium indicators

DMSO, anhydrous

Dimethylsufoxide, anhydrous

EDC (EDAC)

1-(3-dimethylaminopropyl-3-ethylcarbodiimide, hydrochloride)

Flubi-2

Calibration standard for Flubida-2

5 mg

PK-CA707-51024

Ionomycin (calcium salt)

Useful in cell activation experiments when calcium dose-response data are not required (>90% pure)

5 mg

PK-CA577-1566-5

Ionomycin (free acid)

Useful for studies of Ca2+ transport across biological membranes and measurement of cytoplasmic free Ca2+; induces apoptotic neuronal degeneration in embryonic cortical neurons (>90% pure)

5 mg

PK-CA577-1565-5

Pluronic F-127

Nonionic detergent useful for solubilizing large dye molecules such as Fura-2/AM, Fluo-3/AM, Indo-1/AM, and Rhod-2/AM to facilitate cell loading

1g

PK-CA707-59000



30 ml

PK-CA707-59005



1 ml

PK-CA707-59004

Note: Please check our website regularly for new intracellular indicators.

Size

Catalog Number

1 mg

PK-CA707-59006

1 mg 10 mg

PK-CA707-59001 PK-CA577-1501

2 x 50 ml

PK-CA707-59100

10 ml

PK-CA707-90082

100 mg

PK-CA707-59002

6 Cell Analysis

119

6.8 Cell Signaling Assays

Quantitative determination of cAMP/cGMP levels Cell signaling and cell growth/ differentiation studies b-Secretase activity detection Alzheimer’s disease drug development

Features Fast and simple assays Highly sensitive

cAMP and cGMP Immunoassay Kits Adenosine 3’,5’-cyclic monophosphate (cyclic AMP, cAMP) is one of the most important second messengers involved as a modulator in physiological processes. It regulates neuronal, glandular, and cardiovascular processes as well as immune mechanisms, the nervous system, cell growth, and cell differentiation. A number of hormones (e.g. ACTH, TSH, LH) are known to activate cAMP via the action of the enzyme adenylate cyclase which converts ATP to cAMP.

Guanosine 3,5-cyclic monophosphate (cyclic GMP, cGMP) has been shown to be present at levels typically 10-100 fold lower than cAMP in most tissues and is formed by the action of the enzyme guanylate cyclase on GTP. Some hormones (e.g., acetylcholine, insulin, and oxytocin) and chemicals (e.g., serotonin and histamine) cause an increase in cGMP levels. The PromoKine cAMP and cGMP competitive immunoassays were developed for the quantitative determination of cAMP and cGMP levels. They utilize polyclonal capture antibodies against cAMP or cGMP coated on a 96-well plate and HRP-conjugated cAMP or cGMP-HRP which competes with the cAMP or cGMP from the sample. After HRP-substrate addition, the amount of cAMP-HRP or cGMP-HRP bound to the plate can easily be determined spectrophotometrically at 450 nm. The intensity of the blue color is inversely proportional to the cAMP or cGMP concentration in the sample. Due to a novel acetylation procedure 1-100 fmol cAMP or 0.1-10

pmol cGMP can be detected. This is approximately 10 times more sensitive than similar kits.

b-Secretase Fluorometric Assay Kit b-Secretase (BACE) is a membranebound aspartyl protease that cleaves the amyloid precursor protein and is consequently an excellent target for anti-amyloid therapy in the treatment of Alzheimer’s disease. Finding inhibitors of b-secretase is one of the major goals of Alzheimer’s disease drug development. The PromoKine b-Secretase Fluorometric Assay provides a convenient, non-radioactive system for detecting b-secretase activity in biological samples. The kit provides active bsecretase as positive control, b-secretase inhibitor as negative control, optimized peptide substrate (conjugated to two reporter molecules), and buffers for convenient measurement of b-secretase activity in mammalian samples. b-Secretase inhibitors are also available separately.


Size

Catalog Number

cAMP Immunoassay Kit

100 assays

PK-CA577-K371

cGMP Immunoassay Kit

100 assays

PK-CA577-K372

Beta-Secretase Fluorometric Assay Kit

100 assays

PK-CA577-K360

Beta-Secretase Inhibitor I

1 mg

PK-CA577-7501-1

Beta-Secretase Inhibitor II

1 mg

PK-CA577-7502-1


Cell Analysis

Applications

120

6 Cell Analysis

6.9 Cell Stress and Metabolism Kits

Cell Analysis

Applications GST activity measurement and quantitation of GST-tagged fusion proteins SOD assays Determination of the concentration of NO metabolites (nitrate/nitrite) HAT activity measurement Metabolic studies and analysis of mutagenic, carcinogenic, and toxic effects DNA damage detection and quantification Analysis of the cell redox state

CDNB is totally dependent on the presence of active GST. The detection limit is approximately 10 ng active GST/assay. The free form of MCB is almost nonfluorescent, whereas the dye fluoresces blue (Ex/Em = 380/461 nm) when reacting with glutathione. GST catalyzes the MCB-glutathione reaction and the fluorescence levels are proportional to the amount of GST present in the reaction. Therefore, the GST level in samples can be easily detected using a fluorometer or a 96-well fluorometric plate reader.

fluorometric reagent DAN followed by NaOH which converts nitrite into a fluorescent compound (Fluorometric Nitric Oxide Kits) or the addition of Griess Reagent which converts nitrite into a deep purple azo compound (Colorimetric Nitric Oxide Kits). Measurement of the fluorescent compound reaction product or azo chromophore accurately determines the total nitric oxide production. The Fluorometric Assay Kit has been validated with culture media, plasma, and tissue homogenates.

SOD Assay Kit

HAT Activity Assay Kit

Superoxide dismutase (SOD) is one of the most important antioxidative enzymes. The sensitive SOD Assay Kit utilizes WST-1 that produces a watersoluble formazan dye upon reduction with the superoxide anion. The rate of the reduction with a superoxide anion is linearly related to the xanthine oxidase (XOD) activity and is inhibited by SOD (Fig. 8). Therefore, the inhibition activity of SOD can be determined by a colorimetric method.

Histone acetyltransferases (HATs) have been implicated in playing a crucial role in various cellular functions such as gene transcription, cell differentiation, and proliferation. The colorimetric HAT Activity Assay Kit offers a convenient non-radioactive system for rapid and sensitive spectrophotometric detection of HAT activity in mammalian samples. The detection can be continuous and is suitable for kinetic studies.

Fluorometric and Colorimetric Nitric Oxide Kits

DNA Damage Detection Kit

Features Highly sensitive and reproducible detection/measurement Ready-to-use, convenient assays

Fluorometric and Colorimetric GST Assay Kits Glutathione (GSH) is the major intracellular low-molecular-weight thiol and plays a critical role in cellular defense against oxidative stress in tissues and cells. Glutathione S-transferase (GST) is involved in detoxification of xenobiotics mediating conjugation of glutathione with those compounds. The Fluorometric and Colorimetric GST Assays can detect GST activity in crude cell lysates or purified protein fractions and the kits also quantitate GST-tagged fusion proteins. They are based upon the GST-catalyzed reaction between GSH and the GST substrate CDNB (Colorimetric Kit) or MCB (Fluorometric Kit). CDNB has the broadest range of isozyme detectability (a-, µ-, f-, and other GST isoforms). Under certain conditions, the interaction between glutathione and

Nitric oxide (NO) plays an important role in neurotransmission, vascular regulation, immune responses, and apoptosis. The Fluorometric and Colorimetric Nitric Oxide Assay Kits provide an accurate and convenient tool for total nitrate/nitrite concentration determination in a simple two-step process that can be used as a quantitative measure of NO production. The first step is the conversion of nitrate to nitrite utilizing nitrate reductase. The second step involves the addition of the

-

Xanthine O2

2O2

SO3

-

O3S

I N N H

NO2

Xanthine oxidase

-

Uric acid H2O2

WST-1 Formazan

N N

-.

2O2

SO3

-

O3S

I N

N

+ N N

Na + NO2

SOD

O2 + H2O2

Inhibition reaction

Fig. 8: Principle of the SOD Assay Kit.

WST-1

Different physiological and environmental factors such as chemically induced damage and cell aging cause DNA lesions. AP sites (apurinic/apyrimidinic sites) are one of the major types of DNA lesions formed during the process of base excision and repair of oxidized, deaminated, or alkylated bases. It has been estimated that about 2x105 base lesions are generated per cell per day. For this reason, the level of AP sites in cells can be a good indicator of DNA lesions and repairs. The PromoKine DNA Damage Detection Kit utilizes the ARP (Aldehyde Reactive Probe) reagent that reacts specifically with the aldehyde group in the open ring form of the AP sites. After treating DNA containing AP sites with ARP reagents, the AP sites are tagged with biotin residues, which can be quantified using an avidin-biotin assay followed by colorimetric detection. The kit provides the necessary reagents for convenient determination of abasic sites in purified DNA samples in a 96-well plate format.

NAD/NADH and NADP/NADPH Quantitation PromoKine’s NAD/NADH and NADP/ NADPH Quantitation Kits provide a convenient tool for sensitive detection of the intracellular nucleotides NAD, NADH, NADP, and NADPH along with the determination of their ratios. They can be used for studies of energy transformation and the determination of the redox state of cells or tissue. The kits have been designed to specifically recognize NAD/NADH only or NADP/ NADPH only in an enzyme cycling reaction that significantly increases detection sensitivity. Purification of NAD/NADH or NADP/NADPH from sample mixes is not required.

Fluorometric & Colorimetric ATP Quantitation Kit ATP that is formed exclusively in the mitochondria is the major energy currency of living systems and virtually all energy requiring processes utilize the chemical energy stored in the phosphate bond

of ATP. A variety of genetic diseases can affect ATP formation in the mitochondria. There are a number of commercially available ATP assays available that can detect very low amounts of ATP by measuring luminescence (PromoKine Kit PK-CA577-254-200, for example). However, these kits require specialized luminescence instrumentation and utilize luciferase which can be difficult to maintain in an active form. PromoKine’s new Colorimetric & Fluorometric ATP Quantitation Kit is designed to be robust and has a simple straightforward method which utilizes the phosphorylation of glycerol to generate a product that is easily quantified by colorimetric (λmax = 570 nm) or fluorometric (Ex/Em = 535/587 nm) methods. The assay can detect as low as 50 picomole (1 µM) of ATP in various samples.

Fluorometric & Colorimetric ADP Quantitation Kit ADP is a product of ATP dephosphorylation and can be rephosphorylated back to ATP. Dephosphorylation and rephos-

121

phorylation occur via various phosphatases, phosphorylases and kinases. ADP is stored in platelets and can be released to interact with a variety of purinergic receptors. ADP levels regulate several enzymes involved in intermediary metabolism. ADP conversion to ATP primarily occurs within the mitochondrion and chloroplast, although several such processes occur in the cytoplasm. Conventionally, ADP levels have been measured by luciferase/luciferin mediated assays after ADP is converted to ATP. However, the luciferase system is unstable and luminescence equipment is not generally available in most laboratories. PromoKine’s new Fluorometric & Colorimetric ADP Quantitation Kit provides a convenient colorimetric and fluorometric method to measure the ADP level. In this assay, ADP is converted to ATP and pyruvate. The generated pyruvate can be quantified colorimetrically (λmax = 570 nm) or fluorometrically (Ex/Em 535/587 nm). The assay is simple, sensitive, stable, and high-throughput adaptable. It can detect as low as 1 µM ADP in biological samples.

https://www.promokine.info/shop Product Colorimetric GST Assay Kit

Size

Catalog Number

100 assays

PK-CA577-K263

Fluorometric GST Assay Kit

100 assays

PK-CA577-K260


100 assays

PK-CA577-K332

Colorimetric Nitric Oxide Assay Kit

2 x 96 assays

PK-CA577-K262

Fluorometric Nitric Oxide Assay Kit

2 x 96 assays

PK-CA577-K252

SOD Assay Kit DNA Damage Detection Kit

100 assays

PK-CA577-K335

25 assays

PK-CA577-K253

NAD/NADH Quantitation Kit

100 assays

PK-CA577-K337

NADP/NADPH Quantitation Kit

100 assays

PK-CA577-K347

Fluorometric & Colorimetric ATP Quantitation Kit

100 assays

PK-CA577-K354

Fluorometric & Colorimetric ADP Quantitation Kit

100 assays

PK-CA577-K355

100 µg

PK-RP577-1137-100

1g

PK-CA577-1556-1

HAT protein (P/CAF, active) GST Inhibitor-2 (ethacrynic acid) Oxidized Glutathione (GSSG)

1g

PK-CA577-1241-1

Reduced Glutathione (GSSG)

1g

PK-CA577-1242-1

GST protein (active) Superoxide Dismutase (SOD, human)


1 mg

PK-RP577-1243-1

100 µg

PK-RP577-4802-100

Cell Analysis

6 Cell Analysis

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6 Cell Analysis

6.10 Cell Fractionation and Protein Extraction Kits Applications

Membrane Protein Extraction Kit

The Membrane Protein Extraction Kit provides an easy and complete protocol for isolating integral membrane proteins efficiently from cultured mammalian cells. The kit was designed to not only extract the total cellular membrane proteins but also to specifically purify the plasma membrane proteins. The procedure offers consistent yield and high purity (>90%). Membrane proteins prepared using the kit can be utilized in a variety of applications such as Western blotting, 2-D gels, enzyme analyses, etc. The entire procedure takes less than 1 hour.

Cellular protein extraction from mammalian cells Integral membrane protein extraction Preparation of highly enriched cellular fractions (mitochondria, cytosol, nucleus) from mammalian cells

Cell Analysis

Features

Convenient and efficient cell fractionation and protein extraction Easy protocols Proteins and cell fractions can be used for many downstream applications

Mammalian Whole Cell Protein Extraction Kit The Mammalian Whole Cell Protein Extraction Kit was optimized for complete extraction of all cellular proteins. It contains an optimized lysis buffer and a protease inhibitor cocktail as well as other reagents for convenient extraction of mammalian proteins from tissues and cells under non-denaturing conditions. The entire procedure takes less than 20 minutes and the proteins can be used for many downstream applications such as gel electrophoresis, Western blotting, etc.

Mitochondria/Cytosol Fractionation Kit

Nuclear/Cytosol Fractionation Kit The Nuclear/Cytosol Fractionation Kit provides a complete system that enables the separation of nuclear and cytoplasmic fractions of mammalian cells and tissues with little or no crosscontamination, maintaining the nuclear and cytoplasmic compartments separate and intact.

The Mitochondria/Cytosol Fractionation Kit allows isolation of a highly enriched mitochondrial fraction from the cytosol of mammalian cells in a simple and staightforward procedure without ultracentrifugation and toxic chemicals.

Cytosol/Particulate Fractionation Kit The Cytosol/Particulate Fractionation Kit provides an easy and convenient system for rapid separation of cytosol and particulate fractions. The separated fractions are compatible with downstream applications such as assaying cellular distributions of small molecules and proteins or examining signal transduction pathways.

Complete Cell Fractionation Kit The Complete Cell Fractionation Kit is designed for serial sample preparation of four distinct protein fractions, including cytosol, particulate, cytoskeleton, and nuclear fractions from one sample. The prepared fractions are suitable for many downstream assays such as twodimensional gel electrophoresis, protein immunoblotting, gel-shift assays, enzyme activity assays, reporter assays, and more.

https://www.promokine.info/shop Product Mammalian Whole Cell Protein Extraction Kit

Size

Catalog Number

500 assays

PK-CA577-K269


50 assays

PK-CA577-K268

Nuclear/Cytosol Fractionation Kit


PK-CA577-K266-25 PK-CA577-K266-100



PK-CA577-K256-25 PK-CA577-K256-100

Cytosol/Particulate Fractionation Kit

50 assays

PK-CA577-K267

Complete Cell Fractionation Kit

50 assays

PK-CA577-K270

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6.11 Protein Size Markers and Protein Gel Stains

FluoProtein Gel Stain is a fast and sensitive fluorescent dye for visualization and quantitation of proteins separated by 1-D or 2-D SDS-PAGE and an excellent, economic alternative to common fluorescent gel stains. FluoProtein Gel Stain normally emits a low fluorescence, but is strongly fluorescent (bright golden color) when bound to proteins. The maximum emission wavelength of protein-bound FluoProtein Gel Stain is near 570 nm. FluoProtein Gel Stain gives exceptional sensitivity and a wide dynamic range for protein detection. FluoProtein Gel Stain comes as a 100x stock solution that is simply diluted with water by the user to its working concentration. The staining procedure is a simple two-step protocol (fix and stain) that can be completed in only 30 minutes. Gels are fixed with ethanol/acetic acid solution prior to staining with FluoProtein Gel Stain solution. A destain step is normally not required. Gels stained with FluoProtein Gel Stain may be directly visualized with a variety of different UV-based fluorescence imaging systems. The bound FluoProtein Gel Stain is easily removed from the protein by immersing the gel in sufficient water, thus it is compatible with subsequent enzymatic digestion and mass spectrometry for proteomic applications.

GelQuick Protein Gel Stain/ Destaining Kit Traditional staining and destaining methods for SDS-PAGE protein gels

are known to take several hours and also leave a high blue background which reduces the sensitivity of band detection.

ing a broad range (10-245 kDa) with ten blue-stained protein bands and two reference bands at 25 kDa (green) and 75 kDa (red).

PromoKine’s GelQuick Gel Stain/Destaining Kit provides a fast and sensitive method for SDS-PAGE gel staining and destaining. The procedure only takes 5 minutes for gel staining and 15 minutes for destaining. After a total of 20 minutes, visualization of as little as 5 ng of protein on a waterclear background can be seen. The kit contains sufficient solutions to stain and destain up to 40 protein mini gels.

Cell Analysis

FluoProtein Gel Stain

Prestained RTU Protein Ladders PromoKine’s Prestained RTU Protein Ladders are very cost-effective and supplied ready-to-use for direct loading onto gels. They can be used for monitoring protein separation during SDS-polyacrylamide gel electrophoresis, estimation of protein sizes, and verification of Western transfer efficiency onto membranes (PVDF, nylon, or nitrocellulose). They show excellent stability with sharp bands and allow easy identification as bands of different colors are used. Each Prestained RTU Protein Ladder is sufficient for 100-200 mini gels. The Prestained RTU Protein Ladder I contains 9 pink-stained protein markers covering the range from 10 to 175 kDa and three blue-stained reference bands at 10, 40 and 90 kDa. The Prestained RTU Protein Ladder II is a three-color protein standard cover-

Fig. 9: PromoKine’s RTU Protein Ladder I (above) and RTU Protein Ladder II (below) separated on different SDS polyacrylamide gels.


Size

Catalog Number

1 ml 10 ml

PK-MB713-004-1 PK-MB713-004-10

For 40 minigels

PK-MB577-K901

Prestained RTU Protein Ladder I

2 x 0.25 ml

PK-MB713-001P

Prestained RTU Protein Ladder II

0.5 ml

PK-MB713-002P

FluoProtein Gel Stain GelQuick Protein Gel Stain/Destaining Kit

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6 Cell Analysis

6.12 Nucleic Acid Isolation Kits Applications

Mitochondrial DNA Isolation Kit

DNA isolation

Fast, simple, and reproducible procedures Optimal yields Highly pure DNA suitable for various molecular genetic applications

The Mitochondrial DNA Extraction Kit provides a convenient tool for isolating mtDNA from a variety of cells and tissues with high yield and purity, without contamination from genomic DNA. The purified mtDNA can be used for a variety of studies such as enzyme manipulation, Southern blotting, cloning, PCR analysis, and amplifications.

Genomic DNA Isolation Kit I

PCR Purification Kit

The Genomic DNA Isolation Kit I provides a simple and convenient procedure for rapid isolation of genomic DNA from mammalian cells and tissue samples with high yield and purity. The novel method requires less than 90 minutes to prepare highly pure genomic DNA that is free of protein and RNA and is suitable for a variety of applications such as PCR, DNA hybridization, cloning, Southern blotting, and array-based experiments.

The PromoKine PCR Purification Kit has been designed for the work-up of PCR reactions (removal of primer dimers, primers, nucleotides, proteins, salt, agarose, ethidium bromide, and other impurities). The preparation is based on a silica-membrane technology for binding DNA in high-salt and elution in low-salt buffer. The kit provides a simple and efficient way to purify linear or circular DNA and is optimized for working with

Cell Analysis

Features

DNA amounts of up to 50 µg. It requires no organic extractions or precipitation and guarantees high and stable recovery rates.

https://www.promokine.info/shop Product Genomic DNA Isolation Kit I Mitochondrial DNA Isolation Kit PCR Purification Kit

Size

Catalog Number

50 assays

PK-MB577-K281

50 isolations

PK-CA577-K280

50 preparations 250 preparations

PK-MB20-300-50 PK-MB20-300-250

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125

6.13 DNA Size Markers and DNA Gel Stains RTU DNA Ladders

PromoKine’s FluoDNA Gel Stain is a nonmutagenic fluorescent reagent that produces instant visualization of DNA bands upon Blue Light or UV illumination of agarose gels. Supplied in 6x DNA Loading Buffer, it is used to prepare DNA samples and markers for loading onto agarose or polyacrylamide gels. FluoDNA Gel Stain is a highly sensitive stain available for detecting double-stranded DNA (dsDNA) and contains three tracking dyes (Bromophenol Blue, Xylene Cyanol FF, and Orange G) for monitoring the DNA migration during the electrophoresis process. FluoDNA Gel Stain is an economic, safe, and non-hazardous alternative to ethidum bromide. It provides high sensitivity, shows a low background, and causes less DNA damage. It can therefore improve efficiency in downstream applications such as gene cloning.

PromoKine’s RTU DNA Ladders are suitable for use as molecular weight standards for agarose gel electrophoresis. They are very cost-effective and supplied ready-to-use, sufficient for 100-200 mini gels. The RTU DNA Ladder I displays 11 sharp bands ranging from 100 to 1,500 base pairs. Two bands (100 and 1,500 base pairs) have an increased intensity and serve as reference points. The ladder comes ready-to-use containing Orange G and Xylene Cyanol FF as tracking dyes. The RTU DNA Ladder II includes 13 fragments ranging from 250 to 10,000 bp with two bands (1,000 and 3,000 base pairs) which have an increased intensity to serve as reference points. It comes ready-to-use containing Bromphenol Blue as tracking dye.

Base Pairs

DNA Mass (ng/5ml)

Base Pairs

Cell Analysis

DNA Mass (ng/5ml)

FluoDNA Gel Stain

Fig. 10: PromoKine’s RTU DNA Ladder I (left) and RTU DNA Ladder II (right) separated on 1.7% and 1.0% TAE agarose gels, respectively.


Size

Catalog Number

RTU DNA Ladder I (100 bp)

0.5 ml

PK-MB713-001N

RTU DNA Ladder II (1000 bp)

0.5 ml

PK-MB713-002N

1 ml

PK-MB713-003

FluoDNA Gel Stain

Apoptosis Apoptosis, or programmed cell death, is a sophisticated process that involves an intricate series of biochemical events. Specific cell death signals are transduced by different biomolecules such as caspases, which initiate an apoptotic cascade. This process culminates in multiple events such as degradation of cellular proteins and chromosomal DNA, mitochondrial disruption, and changes in plasma membrane integrity.

7


3 7

Apoptosis 7.1 DNA Fragmentation Detection Kits . . . . . 129 7.2 Mitochondrial Apoptosis Staining Kit . . . . 130 7.3 Cytochrome c Apoptosis Detection Kit . . . 130 7.4 Glutathione Detection Kits . . . . . . . . . . . . 131 7.5 Annexin V Detection Kits . . . . . . . . . . . . . 132 7.6 Caspase Kits . . . . . . . . . . . . . . . . . . . . . . . 134 7.6.1

Caspase Detection Kits . . . . . . . . . . . . . . . . 134

7.6.2

Caspase Quantification Kits . . . . . . . . . . . . 136

7.7 Cathepsin/Calpain Detection Kits . . . . . . . 138 7.8

Kinase Detection Kits . . . . . . . . . . . . . . . . 139

7.9

Apoptotic and Necrotic Cell Detection . . . 140

7.10 Apoptotic Cell Isolation Kit . . . . . . . . . . . . 141 7.11 Apoptosis Inducers . . . . . . . . . . . . . . . . . . 141

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129

7.1 DNA Fragmentation Detection Kits

Detecting fragmented nucleosomal DNA Screening for apoptotic cells Fluorescence microscopy Flow cytometry Western blotting

Features High sensitivity Rapid detection of apoptotic cells at the single-cell level Compatible with standard lab equipment Internucleosomal DNA fragmentation is a key sign of apoptosis in mammalian cells. PromoKine offers four different kits for analyzing DNA fragmentation which are based on fluorometric or colorimetric assays. Since all of the kits are designed for use with various multiwell formats, they are also suited for automation.

Fluorometric DNA Fragmentation Detection Kits I and II (BrdU) The PromoKine DNA Fragmentation Detection Kits I and II are based on the TUNEL method and identify DNA strand breaks that occur during apoptosis. The kits provide all required components including positive and negative control cells and tissue slides - for convenient detection of DNA fragmentation by fluorescence microscopy or flow cytometry. The TUNEL assay employs a terminal deoxynucleotidyl transferase (TdT) to

add Br-dUTP (bromolated deoxyuridine triphosphate nucleotides) to the fragmented DNA strands. Br-dUTP is more readily incorporated into DNA strand breaks than other larger ligands (e.g., fluorescein, biotin, or digoxigenin). The incorporated Br-dUTP is subsequently labeled with a green- (Kit I) or a redfluorescent (Kit II) anti-BrdU monoclonal antibody resulting in a very bright signal. The labeled DNA can then be observed by fluorescence microscopy or analyzed by flow cytometry. The DNA Fragmentation Kit II works also very well with GFP transfected cells.

Fluorometric DNA Fragmentation Detection Kit III (F-dUTP) The Fluorometric DNA Fragmentation Detection Kit III is a modified TUNEL assay. It provides all reagents required for the specific detection of apoptotic cells in a cell population, in tissue sections, and in fixed cell preps. The kit utilizes terminal deoxynucleotidyl transferase (TdT) to catalyze the incorporation of fluorescein12-dUTP at the free 3’-hydroxyl ends of the fragmented DNA. The fluoresceinlabeled DNA can then be observed by fluorescence microscopy or analyzed by flow cytometry.

Colorimetric DNA Fragmentation Detection Kit (IHC) The ready-to-use TUNEL-based Colorimetric DNA Fragmentation Detection Kit (IHC) has been specifically designed for expedient detection of DNA frag-

mentation in tissue sections and fixed cell preparations. It includes positive and negative control slides and utilizes Br-dUTP, biotin-labeled anti-BrdU antibodies, and HRP-streptavidin conjugate. Immunohistochemical staining results can be analyzed with a microscope.

Apoptotic DNA Ladder Detection Kit I The Apoptotic DNA Ladder Detection Kit I provides an easy and sensitive method for detecting DNA fragmentation in apoptotic cells. DNA preparation is done in less than 90 minutes without the need for extraction or separation columns. Afterwards, the DNA fragments can easily be visualized by agarose gel electrophoresis. This kit allows the efficient recovery of small DNA fragments ensuring high sensitivity. The Apoptotic DNA Ladder Detection Kit IPlus additionally includes a highly sensitive DNA staining dye for improved visualization of DNA fragmentation by agarose gel electrophoresis.

Apoptotic DNA Ladder Detection Kit II Using the Apoptotic DNA Ladder Extraction Kit II allows the selective isolation of DNA ladders without interference of regular genomic DNA. The kit can detect apoptotic DNA ladders from tissues and cells with an apoptotic cell population of as little as 5%. The kit specifically extracts laddered DNA, not genomic DNA.


Size

Catalog Number

Fluorometric DNA Fragmentation Detection Kit I (BrdU)

60 assays

PK-CA577-K401

Fluorometric DNA Fragmentation Detection Kit II (BrdU)

60 assays

PK-CA577-K404

Fluorometric DNA Fragmentation Detection Kit III (F-dUTP)

50 assays

PK-CA577-K402

Colorimetric DNA Fragmentation Detection Kit (IHC)

50 assays

PK-CA577-K403

Apoptotic DNA Ladder Detection Kit I

50 assays

PK-CA577-K120


50 assays

PK-CA577-K130

Apoptotic DNA Ladder Detection Kit II

50 assays

PK-CA577-K170

Plus

Note: Please check our website regularly for new cell-based apoptosis assays.

Apoptosis

Applications

130

7 Apoptosis

7.2 Mitochondrial Apoptosis Staining Kit Applications Screening for early apoptotic cells Fluorescence microscopy Flow cytometry

Apoptosis

Features

Excellent sensitivity due to high fluorescence intensity Simultaneous detection of apoptotic and healthy cells Excitation and emission spectra are compatible with common excitation sources and filter sets

Disruption of mitochondrial transmembrane potential is one of the earliest intracellular events that occurs upon the induction of apoptosis. The Mitochondrial Apoptosis Staining Kit provides a simple, fluorescent-based in vitro assay for detecting the mitochondrial changes in apoptosis, and the kit distinguishes between healthy and apoptotic cells. The assay utilizes the novel cationic dye MitoCaptureTM that fluoresces differently in healthy versus apoptotic cells. In healthy cells, MitoCaptureTM accumulates and aggregates in the mitochondria - fluorescing brightly red. In

apoptotic cells, a shift in mitochondrial transmembrane potential prevents the dye from accumulating in the mitochondria. Instead, it remains in the cytoplasm in its monomer form - fluorescing green. The fluorescent signals can easily be detected by fluorescence microscopy or flow cytometry using common filters and channels (FITC, rhodamine, PI). The assay takes approximately 20 minutes, is highly sensitive, and detects apoptosis reliably in living cells.

https://www.promokine.info/shop Product Mitochondrial Apoptosis Staining Kit

Size

Catalog Number


PK-CA577-K250-25 PK-CA577-K250-100

7.3 Cytochrome c Apoptosis Detection Kit Applications Screening for apoptotic cells Analysis of apoptotic signal cascades

Features Non-radioactive method Easy and fast procedure No toxic chemicals necessary

Cytochrome c has been shown to play an important role in apoptosis. The protein is located between the inner and outer mitochondrial membranes. An apoptotic stimulus triggers the release of cytochrome c into the cytosol where it binds to Apaf-1. The cytochrome c/Apaf-1 complex activates caspase-9 which then activates caspase-3 and other downstream caspases of the apoptotic cascade. The PromoKine Cytochrome c Apoptosis Detection Kit provides an effective means

for detecting cytochrome c translocation from mitochondria into the cytosol during apoptosis. The kit is easy to use and provides unique reagents for separating a highly enriched mitochondria fraction from the cytosol. Cytochrome c released from mitochondria into the cytosol is then detected by Western blotting using the cytochrome c antibody provided in the kit. The procedure is very simple and straightforward. No ultracentrifugation is required and no toxic chemicals are involved.

https://www.promokine.info/shop Product Cytochrome c Apoptosis Detection Kit

Size

Catalog Number

100 assays

PK-CA577-K257


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131

7.4 Glutathione Detection Kits

Screening for apoptotic cells Glutathione detection Quantitation of glutathione reductase activity

Features Non-radioactive, highly sensitive method Easy and fast procedure Compatible with standard lab equipment

Fluorometric Glutathione Detection Kit Diminished glutathione (GSH) levels occur at the early stages of apoptosis. The Fluorometric Glutathione Detection Kit provides a simple in vitro assay for detecting the GSH changes in apoptosis and other pathological processes. The assay utilizes a dye (monochlorobimane, MCB) that has a high affinity for glutathione. The unbound dye is almost nonfluorescent, whereas the dye fluoresces blue (excitation = 380 nm; emission = 461 nm) when bound to reduced or oxidized glutathione. Consequently, the decrease in glutathione level in apoptotic cells can be easily detected using a fluorometer or a 96-well fluorometric plate reader.

Colorimetric Glutathione Detection Kit The Colorimetric Glutathione Detection Kit provides a convenient colorimetric method for analyzing either total glutathione or only the reduced form of glutathione using a microplate reader. The assay utilizes the glutathione recycling system with DTNB and glutathione reductase. GSH is regenerated from GSSG (oxidized glutathione) by glutathione reductase and reacts again with DTNB to produce yellow 2-nitro-5-thiobenzoic acid that can be detected spectrophotometrically at 412 nm (Fig. 1). This recycling reaction dramatically improves the sensitivity of total glutathione detection. The kit can quantify total glutathione from 1-100 ng/well. For detecting lower glutathione concentrations, such as in

DTNB

blood samples, increasing the reaction time significantly improves the signal. The kit can also specifically detect the reduced form of glutathione (GSH) when glutathione reductase is omitted from the reaction mixture.

Colorimetric Glutathione Reductase Activity Kit PromoKine’s Colorimetric Glutathione Reductase Activity Kit was designed for measuring glutathione reductase (GR) activity in biological samples. It is highly sensitive and well suited for high-throughput. The assay is based on the GR-catalysed reduction of GSSG to GSH, which reacts with 5,5′-Dithiobis (2-nitrobenzoic acid) (DTNB) to generate the yellow TNB2-. The kit can detect 0.1 – 40 mU/ml GR in various samples.

2GSH

Glutathione + NADPH reductase 2-Nitro-5-thiobenzoic acid

GSSG

Fig. 1: Principle of the Colorimetric Glutathione Detection Kit.


Size

Catalog Number

Fluorometric Glutathione Detection Kit

100 assays

PK-CA577-K251

Colorimetric Glutathione Detection Kit

100 assays

PK-CA577-K261

Colorimetric Glutathione Reductase Activity Kit

200 assays

PK-CA577-K761


Apoptosis

Applications

132

7 Apoptosis

7.5 Annexin V Detection Kits Applications

Screening for apoptotic cells Differentiating apoptotic and necrotic cells Annexin detection Flow cytometry Fluorescence microscopy

Apoptosis

Features High fluorescence intensity and sensitivity Many different labels and tags available Fast, convenient protocol suited for automation and high-troughput Excitation and emission spectra are compatible with common excitation sources and filter sets In the early stages of apoptosis, changes can be observed on the cell surface. Cells lose their phospholipid membrane asymmetry and expose phosphatidylserine (PS) on the exterior plasma membrane. Annexin V is a Ca2+-dependent, phospholipid-binding protein with high affinity for PS and is very useful for identifying apoptotic cells that expose PS on their surfaces to the extracellular environment. Annexin V may be conjugated to biotin or fluorophores such as FITC and phycoerythrin (PE). It can also

be tagged with EGFP (Enhanced Green Fluorescent Protein). Since Annexin V conjugates and tagged annexins retain their high affinity for PS, they are ideally suited as highly sensitive probes to identify early apoptotic cells and to detect apoptosis much earlier than assays based on DNA fragmentation or loss of membrane integrity. The stained cells can be evaluated by flow cytometry or fluorescence microscopy using common filter sets and flow cytometer channels.

Annexin V Detection Kits The Annexin V Apoptosis Detection Kits include an Annexin V conjugate or tagged Annexin V as well as binding buffer and in some cases Propidium Iodide (PI) for differentiating between apoptotic and dead cells. The kits allow a one-step staining procedure that takes only 10 minutes. In addition, the assay can be performed directly on living cells. Promokine Annexin V Apoptosis Detection KitPlus includes an additional dye, SYTOX green, that is impermeant to living and apoptotic cells, but stains necrotic cells with intense green fluorescence by binding to cellular nucleic acids. After staining a cell population with e.g., fluorescent Annexin V-Cy3 or Annexin V-PE conjugates, and SYTOX Green dye

in the provided binding buffer, apoptotic cells show red fluorescence, necrotic cells show green fluorescence, and live cells show little or no fluorescence. These populations can easily be distinguished by fluorescence microscopy and flow cytometry using the appropriate filters and channels.

Individual Annexin V Reagents PromoKine also offers a wide range of Annexin V reagents separately. For easy staining of early apoptotic cells, Annexin V conjugated to FITC, PE, Cy3, Cy5, or biotin is available, as well as Annexin V tagged with EGFP. The stained cells can be evaluated by flow cytometry or fluorescence microscopy using common filter sets and flow cytometer channels. Biotinylated Annexin V can be detected in conjunction with conventional dye-staining using any streptavidin- or avidin-dye reagents such as PromoFluorstreptavidin conjugates, fluorescein-, PE-, or APC-streptavidin as well as streptavidin/avidin conjugated to peroxidase, alkaline phosphatase (AP), b-galactosidase, etc. Please visit www.promokine.info to find our complete range of individual Annexin V reagents as well as Annexin V antibodies and ELISAs.


Size

Catalog Number

Annexin V-FITC Detection Kit


PK-CA577-K101-25 PK-CA577-K101-100 PK-CA577-K101-400

Annexin V-FITC Detection KitPlus



Annexin V-Cy3 Detection Kit



Annexin V-Cy3 Detection KitPlus






Annexin V-EGFP Detection Kit



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133

Product

Size

Catalog Number

Annexin V-PE Detection Kit



Annexin V-PE Detection KitPlus



Annexin V-PE-Cy5 Detection Kit



Annexin V-Biotin Detection Kit



Annexin V-FITC Reagent


PK-CA577-1001-200 PK-CA577-1001-1000

Annexin V-Cy3 Reagent

200 assays 1,000 assays

PK-CA577-1002-200 PK-CA577-1002-1000



PK-CA577-1013-200 PK-CA577-1013-1000

Annexin V-EGFP Reagent


PK-CA577-1004-200 PK-CA577-1004-1000

Annexin V-PE Reagent


PK-CA577-1014-200 PK-CA577-1014-1000

Annexin V-PE-Cy5 Reagent


PK-CA577-1015-200 PK-CA577-1015-1000

Annexin V-Biotin Reagent

100 μg 1 mg

PK-CA577-1003-100 PK-CA577-1003-1000

Annexin V Unlabeled Reagent

100 μg

PK-CA577-1005-100

1X Annexin V Binding Buffer

100 ml

PK-CA577-1035

10X Annexin V Binding Buffer

100 ml

PK-CA577-1006


Apoptosis


134

7 Apoptosis

Apoptosis

7.6 Caspase Kits Caspases, a family of cysteine aspartic acid proteases, are important factors in the progression of apoptosis and can be activated by various harmful physiological stimuli such as cytochrome c release due to mitochondrial membrane disruption, DNA damage, ER stress, or inducers such as TNF-alpha and FasL. Caspases are normally present in the cell as inactive pro-enzymes. During apoptosis, they are activated by proteolysis forming a proteolytic cascade that results in the cleavage of distinct proteins that are essential for cell viability.

7.6.1 Caspase Detection Kits

Caspase Staining Kits

Applications

The fluorometric Caspase Staining Kits are the ideal tools for detecting activated caspases in situ in living cells. The kits utilize potent caspase inhibitors conjugated to FITC or rhodamine as fluorescent markers. The caspase inhibitors are cell-permeable, nontoxic, and bind specifically and irreversibly to the activated caspases in apoptotic cells. With the FITC or rhodamine labels, activated caspases in apoptotic cells can be directly visualized by fluorescence microscopy or analyzed by using flow cytometry, or a fluorescence plate reader. Caspase inhibitors without FITC or rhodamine labels are also included in the kits as negative controls. The components of the kits (e.g. the Wash Buffer) are also available separately (please see www.promokine.info).

Screening for apoptotic cells Flow cytometry and fluorescence microscopy Caspase detection

Features High fluorescent intensity and sensitivity Highly specific Fast and simple Appropriate for various multiwell formats and automation Excitation and emission spectra are compatible with common excitation sources and filter sets


Size

Catalog Number

Green Multi-Caspase Staining Kit


PK-CA577-K180-25 PK-CA577-K180-100

Green Caspase-2 Staining Kit


PK-CA577-K182-25 PK-CA577-K182-100



PK-CA577-K183-25 PK-CA577-K183-100



PK-CA577-K188-25 PK-CA577-K188-100



PK-CA577-K189-25 PK-CA577-K189-100



PK-CA577-K172-25 PK-CA577-K172-100

Red Multi-Caspase Staining Kit


PK-CA577-K190-25 PK-CA577-K190-100

Red Caspase-3 Staining Kit


PK-CA577-K193-25 PK-CA577-K193-100



PK-CA577-K198-25 PK-CA577-K198-100



PK-CA577-K199-25 PK-CA577-K199-100

Note: Please check our website regularly for new cell-based apoptosis assays. For a complete list of active recombinant caspases, caspase substrates, inhibitors, and antibodies please visit our website www.promokine.info, or see the PromoKine Price List.

7 Apoptosis

Caspase Drug Screening Kits

The PromoKine Caspase Screening Kit (FC) provides a convenient means for detecting the activation of caspases by flow cytometry in living cells. The assay is based on the cleavage of (aspartyl)2Rhodamine 110 (D2R), a reported substrate for members of caspase family proteases. The caspase substrate D2R is nonfluorescent. However, upon cleavage of the substrate by cellular activated caspases, the released rhodamine 110 gives rise to fluorescence that can be measured by flow cytometry (excitation maximum at 488 nm, emission maximum at 530 nm).

Inhibition of caspases has been shown to delay apoptosis, implicating a potential role in drug screening efforts. PromoKine provides a full line of Caspase Drug Screening Kits for effective screening of caspase inhibitors using fluorometric methods. The assays utilize peptide substrates containing consensus cleavage sequences specific for each caspase, labeled with AFC (7-amino4-trifluoromethyl coumarin). Active caspases (provided in the kits) cleave the synthetic substrates to release free

AFC which can then be quantified using a fluorescence plate reader. Compounds to be screened can be directly added to the reaction, and the level of caspase inhibition can then be determined. The assays are simple, straightforward, and can be performed directly in microtiter plates. Assay conditions for each caspase have been optimized to obtain the maximal activity. Apoptosis

Caspase Screening Kit (FC)

135

https://www.promokine.info/shop Product Caspase Screening Kit (FC)

Size

Catalog Number


PK-CA577-K200-25 PK-CA577-K200-100

Caspase-1 Drug Screening Kit

100 assays

PK-CA577-K151-100


100 assays

PK-CA577-K152-100


100 assays

PK-CA577-K153-100


100 assays

PK-CA577-K154-100


100 assays

PK-CA577-K155-100


100 assays

PK-CA577-K156-100


100 assays

PK-CA577-K157-100


100 assays

PK-CA577-K158-100


100 assays

PK-CA577-K159-100


100 assays

PK-CA577-K160-100

Note: Please check our website regularly for new cell-based apoptosis assays. For a complete list of active recombinant caspases, caspase substrates, inhibitors, and antibodies please visit our website www.promokine.info, or see the PromoKine Price List.

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7.6.2 Caspase Quantification Kits Applications Apoptosis screening Quantification of caspase activity

Apoptosis

Features Highly sensitive and specific Fast and convenient Compatible with standard lab equipment Suited for automation

Caspase Fluorometric & Colorimetric Assay Kits With the Caspase Fluorometric & Colorimetric Assay Kits caspase activity in mammalian cells can be quantified by either fluorescence or absorbance in a fast and simple procedure. The kits utilize the caspase substrate (Ac-DEVD)2-R110 that is fluorogenic as well as chromogenic. It has an extinction coefficient that is 10 times higher than that of pNA - making the assay extremely sensitive.

Caspase Fluorometric Assay Kits The Caspase Fluorometric Assay Kits are intended for convenient and sensitive quantification of caspase activity in cell lysates. The kits utilize synthetic peptides which mimic the optimal substrate for distinct caspases and are labeled with 7-amino-4-trifluoromethyl coumarin (AFC). Activated caspases in apoptotic cells specifically cleave their synthetic substrates, resulting in the release of free AFC which is then quantified using a fluorometer. The kits are fast, straight-forward, and adaptable to 96-well plates.

Caspase Colorimetric Assay Kits The Caspase Colorimetric Assay Kits are intended for convenient and sensitive quantification of caspase activity in cell lysates. The kits utilize synthetic peptides which mimic the optimal substrate for distinct caspases and are labeled with p-nitroaniline (pNA). Activated caspases in apoptotic cells specifically cleave their synthetic substrates, resulting in the release of free pNA, which is then quantified using a spectrophotometer. The kits are fast, straightforward, and adaptable to 96-well plates.

Caspase Fluorometric Assay Kits (HTS) The Caspase Fluorometric Assay Kits (HTS) were designed for economic highthroughput applications. They use the fluorogenic and chromogenic substrate (Ac-DEVD)2-R110 and include specific caspase inhibitors as negative controls.

Caspase-3 and Caspase-7 Immunoassay Kits Caspase-3 plays a key role in the initiation of cellular events during the apoptotic process. The PromoKine Caspase-3 and Caspase-7 Immunoassay Kits allow quantification of the specific activity of caspase-3 and -7. Since caspase-3 and -7 share the same target substrate sequence (DEVD), it is difficult to differentiate the cleavage activity contributed by these two caspases in vitro. Consequently, the assays utilize caspase-3 or -7 specific antibodies to capture the activated caspase-3 or -7 in cell lysate. The specific activity of caspase-3 or -7 can then be analyzed using the common caspase-3/7 substrate DEVDAFC. The assay system ensures specific detection of caspase-3 or caspase-7 ac-

tivity in apoptotic samples. Other caspases and non-specific proteases are not detected.

Individual Caspase Reagents In addition to the complete caspase staining and activity kits, PromoKine also provides a variety of high quality, purified active recombinant caspases and pro-caspases from human, mouse and rat, as well as caspase substrates and inhibitors. Optimized assay buffers and other supplementary reagents for setting up custom assays are also available separately. These reagents allow researchers to perform numerous caspase assays economically. Sets of ready-to-use caspase enzymes, caspase substrates, and caspase inhibitors provide more convenience in setting up individual assays to evaluate multiple caspases in the same experiment. PromoKine’s active caspases are very useful for studying enzyme regulation and kinetics, screening caspase inhibitors, determining target substrates, and as positive controls in caspase activity assays and Western blot analyses. The caspase substrates come in readyto-use form for both fluorometric and colorimetric caspase assays. Please visit www.promokine.info or see the PromoKine Price List to find the complete range of PromoKine individual caspase reagents. A range of caspase antibodies and ELISAs can also be found on our website.

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Product

Size

Catalog Number

Caspase-1 Fluorometric Assay Kit

25, 100, 200, 400 assays

PK-CA577-K110-XXX*


25, 100, 200, 400 assays

PK-CA577-K116-XXX*


25, 100, 200, 400 assays

PK-CA577-K105-XXX*


25, 100, 200 assays

PK-CA577-K126-XXX*


25, 100, 200, 400 assays

PK-CA577-K122-XXX*


25, 100, 200, 400 assays

PK-CA577-K114-XXX*


25, 100, 200, 400 assays

PK-CA577-K112-XXX*


25, 100, 200, 400 assays

PK-CA577-K118-XXX*

25, 100 assays

PK-CA577-K124-XXX*

Caspase-10 Fluorometric Assay Kit Caspase-12 Fluorometric Assay Kit

25, 100 assays

PK-CA577-K139-XXX*

Caspase-1 Colorimetric Assay Kit

25, 100, 200, 400 assays

PK-CA577-K111-XXX*


25, 100, 200, 400 assays

PK-CA577-K117-XXX*


25, 100, 200, 400 assays

PK-CA577-K106-XXX*


25, 100, 200 assays

PK-CA577-K127-XXX*


25, 100, 200, 400 assays

PK-CA577-K123-XXX*


25, 100, 200, 400 assays

PK-CA577-K115-XXX*


25, 100, 200, 400 assays

PK-CA577-K113-XXX*


25, 100, 200, 400 assays

PK-CA577-K119-XXX*

25, 100 assays

PK-CA577-K125-XXX*


PK-CA707-30011-1 PK-CA707-30011-2

1 ml 10 ml 100 ml

PK-CA707-30012-1 PK-CA707-30012-2 PK-CA707-30012-3

Caspase-3 Immunoassay Kit

100 assays

PK-CA577-K163


100 assays

PK-CA577-K167

Caspase-10 Colorimetric Assay Kit Caspase-8 Fluorometric & Colorimetric Assay Kit Caspase-8 Fluorometric Assay Kit (HTS)

Note: Please check our website regularly for new cell-based apoptosis assays. For a complete list of active recombinant caspases, caspase substrates, inhibitors, and antibodies please visit our website www.promokine.info or see the PromoKine Price List.

* XXX stands for the respective package size: 25, 100, 200 or 400 tests.

Apoptosis


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7.7 Cathepsin/Calpain Detection Kits Applications Screening for apoptotic cells

Apoptosis

Features

Non-radioactive method Easy and fast procedure Appropriate for various multiwell formats and automation Compatible with standard lab equipment and various multi-well formats

Apoptosis can be mediated by mechanisms other than the well-studied caspase-mediated cleavage cascade. There is growing knowledge that alternative proteolytic enzymes such as the lysosomal cathepsin proteases may initiate or propagate proapoptotic signals, but it is currently unclear how cathepsin mediates apoptosis.

Cathepsin Activity Assay and Cathepsin Inhibitor Screening Kits The Cathepsin Activity Assay Kits are based on fluorometric or colorimetric (Cathepsin G assay) detection and utilize the preferred substrate sequence for each cathepsin, labeled with AFC

(amino-4-trifluoromethylcoumarin), MCA (7-amino-4-methyl coumarin) or pNA (p-nitroaniline). Cell lysates and other samples that contain cathepsins cleave the synthetic substrate to release free AFC, MCA or pNA that can be easily quantified using a fluorometer/photometer or a fluorescence plate reader/ ELISA reader. The cathepsin assays are easy to perform and can be adapted to 96-well plate assays. Assay conditons have been optimized to obtain the maximal activity. The fluorometric Cathepsin D Inhibitor Screening Kit includes active Cathepsin D enzyme and a synthetic Cathepsin D substrate coupled to MCA. It is a simple, straightforward assay that can be adapted to 96-well plate assays and high-throughput screening. The relative efficacy of test inhibitors are compared to the positive control inhibitor, Pepstatin A. A range of Cathepsin antibodies and ELISAs can also be found on our website.

processes (e.g., apoptosis). The Calpain Activity Assay Kit provides a simple and convenient means for analysing calpain activity in apoptotic cells and other samples. The assay is based on fluorometric detection and quantitation of the cleavage of a calpain substrate at 505 nm using a fluorometer or a fluorescence plate reader. The kit contains positive and negative controls. Comparison of the fluorescence intensity from a treated sample with a normal control allows determination of the changes in calpain activity. The fluorometric Calpain-1 Inhibitor Screening Kit includes active Calpain-1 enzyme and a synthetic Calpain-1 substrate. It provides a rapid, simple, sensitive, and reliable test suitable for high-throughput screening of Calpain-1 inhibitors. In the presence of potent Calpain-1 inhibitors, the hydrolyzation of the calpain substrate will be impeded. An inhibitor control is included to compare the efficacy of test inhibitors.

Calpain Activity Assay and Calpain Inhibitor Screening Kits

A range of different Calpain antibodies can also be found on our website.

The activation of calpain is involved in many physiological and pathological https://www.promokine.info/shop

Product Cathepsin B Activity Assay Kit

Size

Catalog Number

100 assays

PK-CA577-K140

Cathepsin D Activity Assay Kit

100 assays

PK-CA577-K143

Cathepsin D Inhibitor Screening Kit

100 assays

PK-CA577-K148

Cathepsin G Activity Assay Kit

100 assays

PK-CA577-K146

Cathepsin H Activity Assay Kit

100 assays

PK-CA577-K145

Cathepsin K Activity Assay Kit

100 assays

PK-CA577-K141

Cathepsin L Activity Assay Kit

100 assays

PK-CA577-K142

Cathepsin S Activity Assay Kit

100 assays

PK-CA577-K144

Calpain Activity Assay Kit

100 assays

PK-CA577-K240

Calpain-1 Inhibitor Screening Kit

100 assays

PK-CA577-K244

Cathepsin B, human (native)

5 µg

PK-RP577-1021

Cathepsin D, human (native)

5 µg

PK-RP577-1022

Cathepsin H, human (native)

5 µg

PK-RP577-1023

5 µg 100 µg

PK-RP577-1135-5 PK-RP577-1135-100

5 µg

PK-RP577-1025

Cathepsin L, human, recombinant Cathepsin S, human (native) Calpain I, human (native) Cathepsin B & L Substrate Z-FR-R110 Cathepsin Substrate Z-FR-AMC

100 µg

PK-RP577-1134

5 mg

PK-CA707-10209

25 mg

PK-CA707-10201

Calpain Inhibitor Z-LLY-FMK

20 µl (10 mM)

PK-CA577-1125-20C

Cathepsin B&L Inhibitor Z-Phe-Phe-FMK

20 µl (10 mM)

PK-CA577-1126-20C

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7.8 Kinase Detection Kits

Screening for apoptotic cells Kinase detection and activity screening

Features Non-radioactive method Easy, accurate, and fast procedure Compatible with standard lab equipment

large numbers of samples. The assay utilizes an N-terminal c-Jun fusion protein bound to Glutathione-Sepharose beads to selectively “pull down” JNK from cell lysate. After a washing step to remove non-specifically bound proteins, the kinase reaction is carried out in the presence of cold ATP. c-Jun phosphorylation is then determined by Western blot analysis using a phospho-c-Jun specific antibody.

JNK Activity Detection Kit The activation of protein kinases, such as JNK and Akt, plays an important role in apoptosis and other physiological and pathological processes. PromoKine provides a novel non-radioactive system for convenient measurement of JNK and Akt activities.

JNK Activity Screening Kit The JNK Activity Screening Kit can screen JNK activity quickly even with

The JNK Activity Detection Kit utilizes a JNK-specific antibody to immunoprecipitate JNK from cell lysates. The activity of the JNK is then detected in a kinase reaction using recombinant c-Jun as substrate. c-Jun phosphorylation is determined by Western blotting using a phospho-c-Jun specific antibody. The kit specifically detects JNK activity; other kinase activities are not detected.

Akt Activity Detection Kit The Akt Activity Detection Kit utilizes an Akt-specific antibody to immunoprecipitate Akt from cell lysates. Akt-specific activity is then detected in a kinase reaction using recombinant GSK-3a as a substrate. Phosphorylation levels of GSK-3a are determined by Western blot analysis using a phospho-GSK-3a specific antibody included in the kit. The kit specifically detects Akt1, Akt2, and Akt3 activities. Other kinase activities are not detected. Active JNK and Akt kinases as well as JNK/Akt substrates, activators, inhibitors, and control cell lysates are also available separately. Please visit www.promokine.info or see the PromoKine Price List to find more active kinases that can be used for cell analysis and apoptosis research. A range of kinase antibodies can also be found on our website.


Size

Catalog Number

JNK Activity Screening Kit

40 assays

PK-CA577-K430

JNK Activity Detection Kit

40 assays

PK-CA577-K431

Akt Activity Detection Kit

40 assays

PK-CA577-K435

1 mg

PK-CA577-1701

Akt Inhibitor Chk2 Inhibitor

1 mg

PK-CA577-1702

c-Jun (1-79)-GST Fusion Protein

100 ug

PK-CA577-7001

GSK-3a-GST Fusion Protein

100 ug

PK-CA577-7003

GSK-3β, human, recombinant

100 ug

PK-CA577-7004

L-JNK Peptide Inhibitor I

50 µl (1 mM)

PK-CA577-7011

L-JNKI1 Negative Control Peptide

50 µl (1 mM)

PK-CA577-7021

JNK Negative Control Cell Lysate

1 mg

PK-CA577-7031

JNK Activated Cell Lysate

1 mg

PK-CA577-7032

Akt Negative Control Cell Lysate

1 mg

PK-CA577-7035

Akt Activated Cell Lysate

1 mg

PK-CA577-7036

Note: Please check our website regularly for new cell-based apoptosis assays. For a complete list of active recombinant kinases, kinase substrates, inhibitors, and antibodies please visit our website www.promokine.info or see the PromoKine Price List.

Apoptosis

Applications

140

7 Apoptosis

7.9 Apoptotic and Necrotic Cell Detection Applications

Simultaneous detection of apoptotic, necrotic, and healthy cells Fluorescence microscopy Flow cytometry

Apoptosis

Features Convenient ready-to-use kit Fast procedure Highly sensitive

Apoptosis and necrosis, the two major processes leading to cell death, show several distinct differences. One characteristic of apoptotic cells is the translocation of phosphatidylserine from the inner to the outer side of the plasma membrane. This allows phagocytic cells to recognize and destroy apoptotic cells but can also be used for identifying apoptotic cells. Necrotic cells, on the other hand, are characterized by a loss of membrane integrity. This allows membrane-impermeable probes to enter and stain them.

Apoptotic/Necrotic Cells Detection Kit The Apoptotic/Necrotic Cells Detection Kit provides convenient quantification of apoptotic (green) and necrotic (red) cells

within the same cell population by flow cytometry or fluorescence microscopy. For tagging apoptotic cells, fluorescein labeled Annexin V is used. Annexin V specifically binds to the phosphatidylserine exposed on the outer membrane of apoptotic cells. In order to mark necrotic cells, Ethidium homodimer III (EthD-III) is utilized. This fluorescent DNA binding substance can enter necrotic cells because of the damaged plasma membrane, but not living or apoptotic cells. Once in the necrotic cell, it binds to cellular DNA and fluoresces red. Due to its significantly higher fluorescence quantum yield, Ethidium homodimer III is superior to propidium iodide or Ethidium homodimer I.

Fig. 2: Apoptotic U937 cells stained with Annexin V-FITC using PromoKine’s Apoptotic/Necrotic/Healthy Cells Detection Kit.

Apoptotic/Necrotic/Healthy Cells Detection Kit The Apoptotic/Necrotic/Healthy Cells Detection Kit provides convenient quantification of apoptotic (green), necrotic (red), and healthy cells (blue) within the same cell population by flow cytometry or fluorescence microscopy. In addition to the fluorescent dyes utilized in the Apoptotic/Necrotic Cells Detection Kit, it also contains a membrane-permeable blue fluorescent DNA stain (Hoechst 33342) for quantifying healthy cells.

Fig. 3: Apoptotic/Necrotic U937 cell stained with Ethidium homodimer III (EthD-III) using PromoKine’s Apoptotic/ Necrotic/Healthy Cells Detection Kit.


Size

Catalog Number

Apoptotic/Necrotic Cells Detection Kit

50 assays

PK-CA707-30017

Apoptotic/Necrotic/Healthy Cells Detection Kit

50 assays

PK-CA707-30018

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141

Applications Isolation of apoptotic cells Removal of dead cells from culture

Features Fast, simple, and efficient Complete ready-to-use kit The PromoKine Apoptotic Cell Isolation Kit provides a simple and efficient means for isolating apoptotic cells or removing

MagBeads adhere to the magnet while normal cells stay in suspension. The bound apoptotic cells can then be released from the MagBeads using the Elution Buffer provided in the kit. The separated apoptotic cells and normal cells can be used in a variety of assays to study apoptotic mechanisms and pathways. The kit has also been successfully used to remove dead cells from normal cultures. A re-usable magnetic separator optimized for the kit is also available.

dead cells from cell culture or tissue preparations using Annexin V magnetic beads (MagBeads). Since Annexin V is a phospholipid-binding protein with high affinity for phosphatidylserine (PS), it binds strongly to the PS that is redistributed from the inner to the outer surface of the plasma membrane during apoptosis. Annexin V-biotin first binds to apoptotic cells. Then, biotin binds to streptavidin-MagBeads, which enables the separation of apoptotic cells from normal cells. The apoptotic cells bound to the

https://www.promokine.info/shop Product Apoptotic Cell Isolation Kit

Size

Catalog Number

30 isolations

PK-CA577-K258

1

PK-CA577-K1999-1

Magnetic Separator (re-usable)

7.11 Apoptosis Inducers Applications Induction of apoptotic processes Ideally suited for use with diverse apoptosis detection systems

Features High quality Ready-to-use

PromoKine offers a select choice of high quality apoptosis inducers in ready-touse form. Some of them are also available in a convenient set. The apoptosis inducers are well-characterized and effective in disrupting mitochondrial transmembrane potential and activating caspases as well as inducing phosphatidyl-serine (PS) exposure, DNA fragmentation, and other apoptotic characteristics. All inducers are ideally suited for use with the

various PromoKine apoptosis detection systems. PromoKine provides a wide range of high quality growth factors, cytokines, ELISAs, and antibodies that are relevant to apoptosis research (see our website).


Size

Catalog Number

50 µl

PK-CA577-1036-50

Actinomycin D

5 mg 50 mg

PK-CA577-1036-5MG PK-CA577-1036-50MG

Anisomycin

10 mg

PK-CA577-1549-10

1 kit

PK-CA577-K121-5

25 mg

PK-CA577-1552-25

5 mg

PK-CA577-1560-5

50 mg 500 mg

PK-CA577-1039-50MG PK-CA577-1039-500MG

Actinomycin D (10 mM)

Apoptosis Inducer Set Betulinic acid Brefeldin A Camptothecin

Apoptosis

7.10 Apoptotic Cell Isolation Kit

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7 Apoptosis

https://www.promokine.info/shop Product Camptothecin (2 mM) Carboplatin

Catalog Number

1 ml

PK-CA577-1039-1

100 mg

PK-CA577-1553-100

Colchicine

1g

PK-CA577-1519

Cycloheximide

1g

PK-CA577-1041-1G

Cycloheximide (100 mM)

1 ml

PK-CA577-1041-1

10 mg 50 mg

PK-CA577-1524-10 PK-CA577-1524-50

Dexamethasone

1g 10 g

PK-CA577-1042-1G PK-CA577-1042-10G

Dexamethasone (10 mM)

1 ml

PK-CA577-1042-1

Daunorubicin.HCl Apoptosis

Size

Doxorubicin.HCl Etoposide Etoposide (10 mM) Genistein

5 mg

PK-CA577-1527-5

25 mg 250 mg

PK-CA577-1043-25MG PK-CA577-1043-250MG

100 µl

PK-CA577-1043-100

10 mg 100 mg

PK-CA577-1533-10 PK-CA577-1533-100


5 mg

PK-CA577-1566-5


5 mg

PK-CA577-1565-5

Mifepristone

50 mg

PK-CA577-1561-50

Mitomycin C

5 mg 10 mg 100 mg

PK-CA578-M1108-5 PK-CA578-M1108-10 PK-CA578-M1108-100

Okadaic acid

25 µg

PK-CA577-1543-25

Phorbol-12-myristate 13-acetate (PMA)

5 mg

PK-CA577-1544-5

5 mg 50 mg

PK-CA577-1568-5 PK-CA577-1568-50

Rosiglitazone

5 mg 100 mg

PK-CA577-1559-5 PK-CA577-1559-100

Staurosporine

100 µg 1 mg

PK-CA577-1048-010 PK-CA577-1048-1

1g

PK-CA577-1551

Rapamycin

Tamoxifen citrate Thapsigargin

1 mg

PK-CA577-1558-1

Tyrphostin AG 1295

5 mg

PK-CA577-1571-5

Tyrphostin AG 490

5 mg

PK-CA577-1570-5

Note: Please check our website regularly for new apoptosis inducers.

Fluorescent Labeling The availability of new fluorophores has dramatically changed the possibilities for the sensitive detection of biomolecules and the analysis of their interactions. Improved fluorescent dyes are now enabling previously impossible studies of cellular structures and processes. PromoKine provides a wide range of highquality fluorophores for easy labeling of diverse biomolecules including proteins, antibodies, and DNA, as well as complete labeling kits and ready-to-use fluorescent conjugates.

8


3 8

Fluorescent Labeling 8.1

PromoFluor Dyes . . . . . . . . . . . . . . . . . . . 145

8.2

PromoFluor Labeling Kits . . . . . . . . . . . . . 148

8.2.1

PromoFluor Biomolecules Labeling Kits . . . . 148

8.2.2

PromoFluor DNA Labeling Kits . . . . . . . . . . 149

8.3

PromoFluor Labeled Biomolecules . . . . . . 150

8.3.1

PromoFluor Labeled Antibodies . . . . . . . . . 150

8.3.2

PromoFluor Labeled Phalloidin . . . . . . . . . . 150

8.3.3

PromoFluor Labeled Nucleotides . . . . . . . . . 151

8.3.4

PromoFluor Labeled Biotin and Streptavidin 152

8.4

Phycobiliprotein Dyes . . . . . . . . . . . . . . . . 152

8.5

PromoFluor Antifade Reagent . . . . . . . . . 153

8 Fluorescent Labeling

145

8.1 PromoFluor Dyes

Labeling biomolecules such as proteins, antibodies, and nucleic acids Fluorescence microscopy Flow cytometry Fluorescent in situ hybridization (FISH) Fluorescence correlation spectroscopy (FCS) DNA and protein microarrays

Features

Strong light absorption and high fluorescence intensity Very photostable Highly fluorescent over a broad pH range (pH 4 - 9) Water-soluble without the need to use organic solvents Excitation and emission spectra are compatible with common excitation sources and filter sets

PromoFluor dyes are powerful fluorophores spanning almost the whole visible and infrared light spectrum (see Table 1). They are superior to many other commonly used fluorescent dyes and well

qualified for a wide range of cell and molecular biology applications. In addition to the standard dyes, specifically designed PromoFluor-LSS dyes offer an extra large Stokes shift, which makes them ideally suited for multicolor labeling and flow cytometry applications. All PromoFluor dyes are provided either as free carboxylic acids or in three reactive variants ready for covalent coupling to biomolecules: amine-reactive NHS-Esters, AminoModified Labels, and thiol-reactive Maleimides.

PromoFluor NHS-Esters PromoFluor NHS-Esters (N-hydroxysuccinimidyl-esters) are intended for covalently labeling biomolecules containing a primary amino group (all peptides/proteins contain at least one amino group at the N-terminus). Moreover, amino groups can also be incorporated in other biomolecules without changing their activity or functionality. The most common examples are 5’-amino-modified DNA oligomers and cDNA containing aminoallyl-dU.

PromoFluor Amino-Modified Labels PromoFluor Amino-Modified Labels are intended for the covalent labeling of biomolecules containing a carboxy group e.g., glycoproteins. The amino group of PromoFluor Amino-Modified Labels can be cross-coupled to carboxy groups through a mild in situ activation.

PromoFluor Maleimides PromoFluor Maleimides are intended for covalently labeling biomolecules containing a thiol group (e.g. proteins). A neutral thiol-reactive label has little influence on the isoelectric point of a protein. In effect, PromoFluor Maleimides have an advantage over amino group labels because they do not interfere with techniques like differential 2D gel electrophoresis. Since most proteins contain only a few thiol functional groups, thiolreactive dyes can be used to selectively modify a protein or peptide containing a thiol group at a specific site. Moreover, these dyes are well suited for labeling oligonucleotides. Maleimides couple to thiol groups under very mild conditions at a pH around 7.

Fig. 1: Left: Cell structures of HeLa cells visualized using primary anti-beta-tubulin antibodies and secondary PromoFluor 488-labeled antibodies (green) as well as primary anti-centromere antibodies and secondary PromoFluor 547-labeled antibodies (red). Middle: Rat cytokeratin (green) detected with primary rabbit anti-cytokeratin antibodies and secondary anti-rabbit IgGs labeled with PromoFluor 488. Actin (red) visualized using mouse anti-actin antibodies and secondary goat anti-mouse IgGs labeled with PromoFluor 520LSS. Right: Solid-state fluorescence of PromoFluor 633.


Applications

146



Name

Absorption Maximum [nm]

Emission Maximum [nm]

Description

Laser Excitation Source*

PromoFluor 350

354

431

Alternative to Alexa Fluor® 350, AMCA, AMCA-X

PromoFluor 405

400

420

Alternative to Alexa Fluor® 405, Pacific Blue, Cascade Blue

Blue diode laser

PromoFluor 415

418

467

Alternative to Pacific Blue, Spectrum Aqua, and Diethylaminocoumarin

Blue diode laser

PromoFluor 488LSS

485

560

Dye with extra large Stokes shift. Ideally suited for multicolor labeling and flow cytometry applications

Argon 488 laser

PromoFluor 488

493

521

Alternative to Fluorescein (FITC), Alexa Fluor® 488 and CyTM2

Argon 488 laser

PromoFluor 488 Premium

490

516

Very photostable alternative to Fluorescein (FITC), Alexa Fluor® 488, CyTM2

Argon 488 laser

PromoFluor 500LSS

500

630


Argon 488 laser

PromoFluor 505

505

530

Alternative to Rhodamine GreenTM

Argon 488 laser

PromoFluor 510LSS

509

590


Argon 514 laser

PromoFluor 514LSS

515

650


Argon 514 laser

PromoFluor 520LSS

520

664


Argon 514 laser

PromoFluor 532

539

561

Alternative to Alexa Fluor® 532, Tetramethylrhodamine (TMR and TRITC)

Nd:YAG laser

PromoFluor 546

551

572

Alternative to Alexa Fluor® 546, Tetramethylrhodamine (TMR and TRITC)

Helium-Neon 543 laser

PromoFluor 555

557

574

Alternative to Alexa Fluor® 555, CyTM3, and Tetramethylrhodamine (TMR)


PromoFluor 590

581

598

Alternative to Texas Red®


PromoFluor 610

610

630

Alternative to Texas Red


PromoFluor 633

637

657

Alternative to Alexa Fluor 633

Helium-Neon 633 laser and 635 diode laser

PromoFluor 647

653

672

Alternative to Alexa Fluor® 647, CyTM5, and Allophycocyanin

Krypton-Argon 647 laser

PromoFluor 670

673

694

Alternative to Alexa Fluor® 680 and CyTM5.5

Helium-Neon laser

®

®

Argon laser or UV-light

PromoFluor 680

690

709

Alternative to Alexa Fluor 680, Cy 5.5

Helium-Neon laser

PromoFluor 700

704

723

Alternative to Alexa Fluor® 700

Far-red diode lasers

PromoFluor 750

748

772

Alternative to Alexa Fluor® 750, CyTM7

Far-red diode lasers

PromoFluor 770

769

796

Alternative to Alexa Fluor® 750 and CyTM7

Xenon-arc lamp or dye-pumped laser

PromoFluor 780

783

800

Near infrared dye allowing for deep tissue penetration and low background


PromoFluor 840

844

-

Near infrared dye allowing for deep tissue penetration and low background


®

TM

Table 1: List of currently available PromoFluor dyes (for new PromoFluor dyes that may be added in the future please visit our website). *PromoFluor dyes are compatible with common excitation sources including diode lasers, LEDs, tungsten lamps, xenon arc lamps, and custom made systems.


147

Excitation / Emission [%] 100 80 60

Fig. 2: Excitation (left) and emission (right) spectra of PromoFluor 488LSS.


40 20

300

350

400

450

500

550

600

650

700

Wavelength [nm]

PromoFluor ChipPacks PromoFluor ChipPacks are pre-packed PromoFluor dyes intended to label

cDNA for microarray assays. The packs contain 20 µg aliquots of PromoFluor 555 and PromoFluor 647 NHS-Ester. These aliquots are sufficient for labeling

the amount of cDNA synthesized in a standard reverse transcription assay with enzymatic incorporation of aminoallyldUTP.


Size

Catalog Number*

PromoFluor Carboxylic Acid

1 mg 5 mg

PK-PFXXX-0-01 PK-PFXXX-0-05

PromoFluor NHS-Ester

1 mg 5 mg


PromoFluor Amino-Modified Label

1 mg 5 mg


0.5 mg 1 mg 5 mg

PF-PFXXX-3-005 PK-PFXXX-3-01 PK-PFXXX-3-05

20 labeling reactions 50 labeling reactions 100 labeling reactions

PK-PF-CP-S PK-PF-CP-M PK-PF-CP-L

PromoFluor Maleimide

PromoFluor ChipPack

Coupling protocols for proteins are available on our website: www.promokine.info Note: Different standard sizes are available for PF-488 and PF-505 carboxylic acids and NHS esters.

* XXX- stands fo the respective PromoFluor dye (e.g. PF-350, PF-405, PF-415 etc., see Table 1). Please see our website for an up-to-date list of available PromoFluor Dyes.

148


8.2 PromoFluor Labeling Kits


8.2.1 PromoFluor Biomolecules Labeling Kits Applications

Features

Labeling biomolecules such as proteins, antibodies, and nucleic acids Fluorescence microscopy Flow cytometry Fluorescent in situ hybridization (FISH) Fluorescence correlation spectroscopy (FCS)

Convenient ready-to-use kits Contain superior PromoFluor dyes (see page 146) Highly efficient labeling Very bright and stable fluorescent signals

PromoFluor Biomolecules Labeling Kits were developed for the convenient and highly efficient labeling of target proteins or antibodies with PromoKine’s superior PromoFluor dyes. They contain the respective PromoFluor NHS-Ester as well as all the other components required for labeling biomolecules. PromoFluor Biomolecules Labeling Kits work with all molecules with a primary amino group such as peptides, proteins, 5’-aminomodified DNA oligomers, and cDNA containing aminoallyl-dU.

Fig. 3: Cell structures labeled with antibodies bound to green-fluorescing PromoFluor 505 (left) and red-fluorescing PromoFluor 647 (red) dyes. Illustrations kindly provided by Covalys Biosciences AG, Switzerland.

https://www.promokine.info/shop Product PromoFluor-XXX-Labeling-Kit

Size

Catalog Number*

5 reactions 10 reactions

PK-PF-LK-XXX-05 PK-PF-LK-XXX-10

*XXX- stands fo the respective PromoFluor dye (PF-405, PF-415, PF-488P, PF-488LSS, PF-514LSS, PF-546, PF-555, PF-633, PF-647, PF-670, PF-750, PF-770; please see our website for an up-to-date list of available PromoFluor Labeling Kits).


149

Applications

DNA labeling by PCR DNA labeling by nick translation Labeling of probes for FISH, microarrays, Southern hybridizations, etc.

Features Convenient and fast DNA labeling Optimal incorporation of labeled dUTPs Highly fluorescent and stable fluorophores PromoKine’s PCR Labeling Kits and Nick Translation Labeling Kits were designed for convenient direct DNA labeling by PCR or nick translation using Taq polymerase or DNA Polymerase I/DNase I. The fluorescently labeled dUTP analogs provided with the kits have been optimized for enzymatic incorporation into DNA by using a proprietary linker technology. The high incorporation rates

of the dye-dUTP analogs combined with the outstanding stability and quantum yield of the fluorophores make the kits the ideal choice for all typical DNA labeling applications such as FISH, single molecule detection, microarray gene expression profiling, and other nucleic acid hybridization assays. PromoKine’s PCR Labeling Kits contain all reagents required for highly efficient, rapid, and easy-to-perform DNA labeling (excluding primers and template DNA). The Nick Translation Labeling Kits contain all reagents required for efficient, rapid and easy-to-perform DNA labeling by nick translation (excluding template DNA). The labeled dUTPs are also available separately (see page 151). In addition, we also provide a kit for nonfluorescent labeling of DNA with biotin. The biotinylated DNA can then interact with streptavidin or strepatavidin conjugates (see page 152) for detection or purification applications.


8.2.2 PromoFluor DNA Labeling Kits

Fig 4: PromoKine’s DNA Labeling Kits have been used to generate fluorescently labeled DNA probes for staining specific chromosome regions.


Product Description

Size

Catalog Number

PromoFluor-430 PCR Labeling Kit

Blue-green fluorescent DNA labeling by PCR


PK-PF430-PCRLK-10 PK-PF430-PCRLK-50


Green-fluorescent DNA labeling by PCR




Yellow-fluorescent DNA labeling by PCR




Orange-fluorescent DNA labeling by PCR




Red-fluorescent DNA labeling by PCR



TexasRed PCR Labeling Kit

Orange-fluorescent DNA labeling by PCR


PK-PFTXR-PCRLK-10 PK-PFTXR-PCRLK-50

Biotin PCR Labeling Kit

Kit for biotin-labeling of DNA by PCR


PK-CA-BIO-LK100 PK-CA-BIO-LK500

PromoFluor-430 Nick Translation Labeling Kit

Blue-green fluorescent DNA labeling by nick translation


PK-PF430-NTLK-10 PK-PF430-NTLK-50


Green-fluorescent DNA labeling by nick translation




Yellow-fluorescent DNA labeling by nick translation




Orange-fluorescent DNA labeling by nick translation




Red-fluorescent DNA labeling by nick translation



Note: Please see our website for an up-to-date list of available PromoFluor DNA Labeling Kits.

150


8.3 PromoFluor Labeled Biomolecules 8.3.1 PromoFluor Labeled Antibodies Applications


Fluorescence microscopy Flow cytometry Fluorescent in situ hybridization (FISH) Fluorescence correlation spectroscopy (FCS)

Features

Ready-to-use antibody conjugates of all PromoFluor dyes (see page 146) are . available as goat anti-human IgGs, goat anti-rabbit IgGs, and goat anti-mouse IgGs. The labeled secondary antibodies (polyclonal) are affinity-purified and dissolved in 100 mM PBS (pH 7.5) at a concentration of 1 mg/ml. Each antibody comes with a batch-specific absorption spectrum. Fig 5: Tubulin and centromere structures of HeLa cells visualized by using specific primary antibodies and secondary antibodies labeled with PromoFluor 488 and PromoFluor 555.

High quality and ready-to-use Labeled with PromoFluor dyes High intensity yields and stable fluorescent signals


Size

Catalog Number*

PromoFluor-XXX-Goat Anti-Mouse IgG

1 mg

PK-PFXXX-AK-M1

PromoFluor-XXX-Goat Anti-Rabbit IgG

1mg

PK-PFXXX-AK-R1

PromoFluor-XXX-Goat Anti-Human IgG

1 mg

PK-PFXXX-AK-H1

*XXX- stands for the respective PromoFluor dye (PF405, PF488P, PF488LSS, PF500LSS, PF505 etc). Please see page 146 for details.

8.3.2 PromoFluor Labeled Phalloidin Applications Localization and visualization of actin filaments Fluorescence microscopy

Features

High quality and ready-to-use Labeled with the superior PromoFluor dyes (see page 146) Yields highly intensive and stable fluorescent signals

Phalloidin binds very selectively to actin (much more tightly to actin filaments than to actin monomers). In combination with conjugated PromoFluor dyes, phalloidin is enormously useful in localizing actin filaments in living or fixed cells as well as for visualizing individual actin filaments in vitro. Ready-to-use phalloidin conjugates of all PromoFluor dyes are available (10 nmol, 300 units). Fig. 6: Phalloidin and centromere structures of HeLa cells stained using a PromoFluor 647-Phalloidin conjugate and primary anti-centromer antibodies in combination with secondary antibodies labeled with PromoFluor 488.

https://www.promokine.info/shop Product PromoFluor-XXX, phalloidin conjugate

Size*

Catalog Number**

10 nmol

PK-PFXXX-7-01

*10 nmol are sufficient for staining approximately 300 microscopic slides. **XXX- stands for the respective PromoFluor dye (PF405, PF488P, PF488LSS, PF500LSS, PF505 etc). Please see page 146 for details.


151

Applications

Features

properties with regard to signal intensity, water solubility, and stability. In addition, the low molecular weight of the dyes results in minimal steric hindrance and unproblematic incorporation into DNA using conventional enzymatic techniques like PCR/RT-PCR. AminoallyldUTP can be incorporated into DNA by Taq polymerase, Klenow fragment, and reverse transcriptases. All PromoFluor labeled nucleotides are delivered as 1 mM ready-to-use aqueous solutions.

Fluorescent labeling of nucleic acids Fluorescent in situ hybridization (FISH) Multicolor FISH (chromosome painting) Microarrays

High quality and ready-to-use Labeled with superior PromoFluor dyes (see page 146) High intensity yields and stable fluorescent signals Minimal steric hindrance PromoKine provides aminoallyl-dUTP (deoxyuridine-triphosphate) conjugates for all PromoFluor dyes (see page 146). The labeled nucleotides offer excellent

Fig 7: 2-color labeling of chromosomes 1 and 8 using dUTPs conjugated to PromoFluor 505 and PromoFluor 555. Picture kindly provided by Applied Spectral Imaging.

Fig 8: Fluorescence laser scan of a microarray slide using PromoFluor dyes. Different matching oligonucleotides, negative controls, and reference oligonucleotides were spotted in 25 replicates and at 4 different concentrations to epoxysilane coated slides and hybridized with PromoFluor labeled samples. Picture kindly provided by the Functional Genomics Center Zurich, Switzerland. https://www.promokine.info/shop

Product PromoFluor-XXX-aadUTP

Size

Catalog Number*

100 µl

PK-PFXXX-8-100

PromoFluor-430-dUTP (NT)

10 µl 50 µl

PK-PF430-DUTP-10N PK-PF430-DUTP-50N

PromoFluor-430-dUTP (PCR)

10 µl 50 µl

PK-PF430-DUTP-10P PK-PF430-DUTP-50P


10 µl 50 µl



10 µl 50 µl



10 µl 50 µl



10 µl 50 µl



10 µl 50 µl



10 µl 50 µl



10 µl 50 µl



10 µl 50 µl


TexasRed-dUTP (PCR)

10 µl 50 µl

PK-PFTXR-DUTP-10P PK-PFTXR-DUTP-50P

Biotin-dATP (PCR)

10 µl (1 mM) 50 µl (1 mM)

PK-CA-BIO-DATP-10P PK-CA-BIO-DATP-50P

Biotin-dCTP (PCR)

200 µl (1 mM) 1 ml (1 mM)

PK-CA-BIO-DCTP-200P PK-CA-BIO-DCTP-1000P

Biotin-dUTP (PCR)

200 µl (1 mM) 1 ml (1 mM)

PK-CA-BIO-DUTP-200P PK-CA-BIO-DUTP-1000P

* XXX- stands for the respective PromoFluor dye (PF-405, PF-488P, PF-488LSS, PF-500LSS, PF-505 etc). Please see page 146 for details.


8.3.3 PromoFluor Labeled Nucleotides

152


8.3.4 PromoFluor Labeled Biotin and Streptavidin Applications Fluorescence microscopy Flow cytometry Fluorescent in situ hybridization (FISH) Fluorescence correlation spectroscopy (FCS)


Features

High quality and ready-to-use Labeled with superior PromoFluor dyes (see page 146) High intensity yields and stable fluorescent signals

Streptavidin has a very strong affinity for biotin with a dissociation constant of approximately 10-15 mol/l. This is

the highest known affinity between any protein and its ligand. Extensive chemical modifications have little effect on this affinity. Streptavidin is a tetrameric 53 kDa protein. Since it lacks any carbohydrate modification and has a near-neutral pH, it has the advantage of much lower nonspecific binding than avidin. On the other hand, streptavidin is less watersoluble than avidin. Ready-to-use streptavidin and biotin conjugates of all PromoFluor dyes (see page 146) are available. The streptavidin conjugates are affinity-purified and dissolved in 100 mM PBS (pH 7.5) at a concentration of 1 mg/ml. The biotin conjugates are affinity-purified and lyophilized. Each conjugate comes with a batch-specific absorption spectrum.

Fig. 9: Staining of plasma membranebound neurokinin-1 receptor (red) in a U2OS cell using a PromoFluor 547-conjugate. Picture kindly provided by Covalys Biosciences AG, Switzerland.


Size

PromoFluor-XXX, streptavidin conjugate PromoFluor-XXX, biotin conjugate

Catalog Number*

1 mg in 1 ml PBS

PK-PFXXX-5-01

1 mg

PK-PFXXX-6-01

Note: Please see our website www.promokine.info for an up-to-date list of available fluorescent labeled biomolecules.

*XXX- stands for the respective PromoFluor dye (PF-405, PF-488P, PF-488LSS, PF-500LSS, PF-505 etc). Please see page 146 for details.

8.4 Phycobiliprotein Dyes Phycobiliproteins are water-soluble proteins with covalently bound chromophores present in cyanobacteria and certain algae that capture light energy during photosynthesis. The major phycobiliproteins are R-Phycoerythrin (R-PE), B-Phycoerythrin (B-PE), and Allophycocyanin (APC).

as large Stokes shifts. The fluorescence yield is equivalent to at least 30 unquenched fluorescein or 100 rhodamine molecules at comparable wavelengths (on a molar basis), and fluorescence is not quenched by external agents since it is protected by covalent binding to the protein backbone.

Phycobiliproteins can be used as fluorescent dyes to label biomolecules to obtain specific probes or markers for highly sensitive cell and molecular biological applications such as immunofluorescence microscopy, flow cytometry, microarrays, FISH, enzyme kinetic studies, etc. They show optimal quantum yields (up to 98%) and extinction coefficients as well

PromoKine’s phycobiliproteins can easily be linked to antibodies. The sensitivity of antibodies conjugated with R-PE and B-PE is usually five to ten times greater than that of the corresponding fluorescein conjugate. PromoKine offers R-PE, B-PE, and APC for labeling of biomolecules such as pro-

teins, antibodies, and nucleic acids. They show strong light absorption, high fluorescence yields, excellent stability and high fluorescence over a broad pH range and are compatible with common excitation sources and filter sets. Please see www.promokine.info/products/fluorescent-labeling for more information and pricing.


153

Applications Mounting medium for fluorescence microscopy

Features

Significantly increased photostability Compatible with many common dyes Conserves cell preparations Convenient packaging

Irreversible photobleaching processes lead to a loss of fluorescent signal sensitivity particularly when high intensity excitation light is used for a longer duration and the target molecules in samples are in low abundance. PromoFluor Antifade Reagent significantly increases the photostability of fluorochromes in fixed cells, fixed tissues, and cell-free preparations. It is compatible with many common dyes and dye complexes resulting in prolonged signals and improved image con-

trast especially in multicolor applications. Moreover, it gently solidifies after the attachment of the cover slide, permanently conserving the cell preparation and fixing the coverslip. PromoFluor Anitfade Reagent makes slide immobilization using e.g. nail polish unnecessary. Samples can be saved for months after mounting, retaining fluorescence during prolonged storage. PromoFluor Antifade Reagent comes premixed in a convenient dropper bottle.

Fig. 10: Immunofluorescence imaging of a human keratinocyte stained with FITC-labeled antibodies without (top) and with (bottom) PromoFluor Antifade Reagent at different excitation intervals (5, 10, 15, 20, 25 seconds).


Product Description

Size

Catalog Number

PromoFluor Antifade Reagent

Mounting Medium for Fluorescence Microscopy

8 ml

PK-PF-AFR1


8.5 PromoFluor Antifade Reagent

Cell Transfection The ability to introduce foreign nucleic acids into eukaryotic cells has become a powerful tool in studying and controlling gene expression. Cloned genes can be transfected into cells for biochemical characterization, mutational analyses, investigating gene regulatory elements, and producing specific proteins. Cell transfection promotes the rapid advancement of our knowledge of genetic regulation and protein function.

9


3 9

Cell Transfection 9.1 PromoFectin Transfection Reagents . . . . . 157 9.2 Magnet Assisted Transfection (MATra) . . . 161 9.3 Transfection Accessories . . . . . . . . . . . . . . 165 9.3.1 GeneBooster . . . . . . . . . . . . . . . . . . . . . . . . 165 9.3.2

9.4

Cell Clean . . . . . . . . . . . . . . . . . . . . . . . . . . 165

Reporter, Expression and Fusion Vectors . . 166

9.4.1

pPK-CMV Reporter Vectors . . . . . . . . . . . . 166

9.4.2

pPK-CMV Expression Vectors . . . . . . . . . . . 166

9.4.3

pPK-CMV Fusion Vectors . . . . . . . . . . . . . . 166

9 Cell Transfection

157

9.1 PromoFectin Transfection Reagents PromoFectin

PromoFectin is a series of reagents for highly efficient and reproducible transfection of a variety of adherent and non-adherent cells that can be used to transfect nucleic acids or proteins. PromoFectin comes either as a broadrange reagent or as specialized reagents for specific cell types and biomolecules.

Efficient transfection of many cell types including primary cells Suited for plasmids, oligonucleotides, siRNA, and polypeptides For stable and transient transfections

Features Superior transfection efficiency Fast procedure and easy handling without medium change Compatible with serum and antibiotics Good cell survival and viability after transfection

The standard broad-range PromoFectin is a non-liposomal transfection reagent based on a water-soluble, cationic polymer. It is free of animal-derived components and shows very low cytotoxicity. PromoFectin ensures highly effective delivery of plasmids, oligonucleotides, and siRNA into a variety of adherent

Cytoplasm

Nucleus

Nucleic acid

PromoFectin

Cationic complexes

Endocytosis

Endosome

Proton sponge

Nucleic acid in 150 mM NaCl solution

Incubate 30 minutes at RT PromoFectin

Incubate 24-28 h. Then, measure protein expression.

and non-adherent cell types with a high degree of reproducibility. Since its development, PromoFectin has been used successfully with many problematic cell lines and sensitive primary cells.

Cell Transfection

Applications

Fig. 1: Primary fibroblasts (NHDF, see page 27) transfected with the GFP reporter vector pPK-CMV-R4 (see page 166) using PromoFectin.

Fig. 2: PromoFectin assembles nucleic acids into compact structures that bind to the cell surface and are delivered into the cell by endocytosis. In the endosome, PromoFectin acts as a proton sponge, buffering the endosomal pH that leads to pH inhibition of endosomal nucleases. Then, continuous proton influx leads to osmotic swelling and rupture of the endosome - allowing for almost complete release of the nucleic acids into the cytosol. Moreover, PromoFectin also favours DNA trafficking to the cell nucleus – a property that is particularly useful for transfecting slow-dividing or resting cells.

Fig. 3: The PromoFectin transfection procedure is fast and easy. Add PromoFectin to the nucleic acid solution, mix, incubate for 15-30 minutes, add growth medium (which can contain serum and antibiotics), and pipette the complexes onto the cells. The cells are then incubated for further analysis. No post-transfection removal of complexes or medium change/addition is necessary.

158

9 Cell Transfection

Cell Transfection

Fig. 4: Comparison of transfection efficiencies for PromoFectin and three other common transfection reagents recommended for primary cells. Primary human fibroblasts (NHDF, see page 27) cultured in 24-well plates were transfected with a luciferase reporter plasmid*. The appropriate protocols recommended by the manufacturers were used for transfection with PromoFectin and the competing reagents. All transfections were performed in triplicate (values normalized to 1 mg protein).

* See page 166 for reporter plasmids and page 107 for reporter assays.

PromoFectin-HUVEC

PromoFectin-Hepatocyte

PromoFectin-Macrophage

PromoFectin-HUVEC is a non-liposomal PromoFectin derivative optimized for the transfection of primary endothelial cells such as HUVEC1 and HDMEC2. It shows very little cytotoxicity and enables very high transfection rates also with fragile, sensitive cell types. Once inside the cell, PromoFectin’s unique “proton sponge” property provides an excellent escape mechanism for DNA particles to the cytoplasm.

PromoFectin-Hepatocyte is a ligandconjugated, non-liposomal derivative of PromoFectin specialized for hepatocyte transfection (e.g., primary human and murine hepatocytes, hepatocyte cell lines, and hepatocarcinoma cells). The reagent/nucleic acid complexes bind specifically to surface receptors on the target cells leading to extremely high transfection rates.

PromoFectin-Macrophage was developed for optimal transfection of the known hard-to-transfect macrophages (e.g. primary human macrophages obtained from blood monocytes) and macrophage-derived cell lines such as RAW 264.7. PromoFectin-Macrophage is a mannose-bearing variant of the standard PromoFectin reagent. It binds to cells expressing mannose-specific membrane receptors - such as macrophages, monocytes and dendritic cells leading to enhanced internalization of the DNA complexes.

Fig. 5: Primary human endothelial cells (HUVECs) transfected with the reporter vector pPK-CMV-R4 (see page 166) using PromoFectin-HUVEC. Nuclei counterstained with DAPI.

Fig. 6: Primary human hepatocytes transfected with the reporter vector pPK-CMV-R4 (see page 166) using PromoFectin-Hepatocyte.

Fig. 7: Human monocyte-derived macrophages were transfected with a galactosidase-expressing plasmid after 7 days of culture in the presence of 100 U/ml GM-CSF and serum-containing medium .

1 2

Human Umbilical Vein Endothelial Cells (see page 13) Human Dermal Microvascular Endothelial Cells (see page 17)

9 Cell Transfection

159

Cell Transfection

Fig. 8: Comparison of transfection efficiencies for PromoFectinHUVEC, PromoFectin, and three other common transfection reagents recommended for primary cells. Primary HUVEC (see page 13) cultured in 24-well plates were transfected with a luciferase reporter plasmid*. The appropriate protocols recommended by the manufacturers were used for transfection with PromoFectin and the competing reagents. All transfections were performed in triplicate (values normalized to 1 mg protein).

* See page 166 for reporter plasmids and page 107 for reporter assays.

PromoFectin-Neuron PromoFectin-Neuron is a cationic polymer-based transfection reagent created for the transfection of primary neuronal cells, glial cells, and neuronal cell lines. Cells take up the polymer/DNA complex through endocytosis and the polymer is biodegraded into non-toxic molecules. Compared to viral methods, PromoFectinNeuron allows less complicated and faster nucleic acid delivery with high transfection rates.

PromoFectin-Insect PromoFectin-Insect is a unique, lipsomebased transfection reagent optimized for nucleic acid delivery into insect cells, especially those derived from Spodoptera frugiperda (e.g. Sf9 or Sf21). It outperforms other insect cell transfec-

Fig. 9: Primary neurons transfected with the reporter vector pPK-CMV-R4 (see page 166) using PromoFectin-Neuron.

tion reagents in comparison studies and requires less DNA. PromoFectin-Insect demonstrates broad vector delivery capability, excellent reproducibility, and high expression levels for small- or largescale protein production.

a wide variety of cell types, including problematic cell lines and primary cells. It enables optimal gene silencing with low cytoxicity.

PromoFectin-CellLine

PromoFectin-Polypeptide is a novel liposomal reagent created for the highly efficient and fast delivery of proteins and polypeptides (e.g. blocking peptides and labeled antibodies) into a wide variety of cells. It captures protein molecules and transports them into the cytosol of living cells while maintaining the protein’s structure and functionality. This allows researchers to study intracellular protein functions without the need for DNA cloning and DNA transfection. PromoFectin-Polypeptide is also available in pre-coated single-use tubes, providing additional convenience and speed.

PromoFectin-CellLine is a cationic lipidbased reagent developed to obtain optimal transfection results with several widely used cell lines (CHO, HeLa, COS, 293, NIH-3T3) at low costs. Its formulation and the provided protocol allow fast and convenient cell transfection.

PromoFectin-siRNA PromoFectin-siRNA is a cationic lipidbased transfection reagent designed for the highly efficient delivery of siRNA into

PromoFectin-Polypeptide

Fig. 10: Left: NIH-3T3 cells transfected with the GFP reporter plasmid pPK-CMV-R4 (see page 166) using PromoFectin-CellLine. Right: CHO cells transfected with the GFP reporter plasmid pPK-CMV-R4 (see page 166) using PromoFectin-CellLine.

160

9 Cell Transfection

Cell Transfection

n Some cell types successfully transfected with PromoFectin

a

Adipocyte cells

Primary human pre-adipocytes

Endothelial-like cells

BBMC; BAEC; HLMECa, HUVECa, H-MVEC, primary human Lung Lymphatic Endothelial cells, primary HUVECa, primary HDMECa

Epithelial-like cells

2CSFMEo, 3LL, 56FHTE8o, 6CSFMEo, 9HTEo-, A549, AsPC-1, BxPC-3, CaCo2, Ca Ski, CFNPE90- CF, CFPEo, CFSMEo, CHO, D 407, DHD Pro.b, HeLa, Huh-7D12, IGROV1, MCF-7, MDA-MB435S, MDCK, MLE-12, MLE-15, MRC-5, PGHAM-1, SAOS-2, SiHa, SKOV3, U-2 OS, HEK293, HEK293FT

Fibroblast-like cells

BHK-21, COS-7, CV-1, L929, NIH 3T3, MEF, primary human skin fibroblasts (NHDF)

Hepatocyte-like cells

BNL, CL 2b, Hepa 1-6, HepG2b, Huh-7b, M-PAC, primary human/mouse/rat hepatocytesb

Keratinocytes

Primary human keratinocytes

Pancreatic carcinoma cells

AR42J, BxPC-3, MIAPaCa-2, PanC-1, PGHAM-1

Leukemia cells / lymphoblasts

Daudi, Jurkat, K562, Molt 4

Melanoma cells / melanocytes

B16-F10, primary human epidermal melanocytese

Myotubes / myoblasts / muscle cells

C2C12

Neuronal cells

CLBPEC, Neuro2A, PC12, SH-SY5Y, SHEP, SK-N-MC, F11, NG 108-15d, primary chicken embryonic neurons, primary rat and mouse cortical neuronsd, primary rat hippocampal neuronsd, primary rat cerebellar granular neuronsd, primary rat striatal neuronsd

Chondrocytes

HCS-2/8

Glioblastoma

U87

Blood cells

Human macrophagesc, monocyte-derived macrophagesc, RAW 264.7c, THP-1c

Mesenchymal stem cell

STRO-1+ DaMSCs cells

PromoFectin-HUVEC,

b

PromoFectin-Hepatocyte,

c

PromoFectin-Macrophage,

d

PromoFectin-Neuron,

e

Promofectin-siRNA

Please contact us or see www.promokine.info for information about new cell types that have been successfully transfected using PromoFectin. https://www.promokine.info/shop Product

Size

Catalog Number

PromoFectin

0.1 ml 0.5 ml 1 ml

PK-CT-2000-10 PK-CT-2000-50 PK-CT-2000-100

PromoFectin-HUVEC

0.1 ml 0.5 ml

PK-CT-2000-HUV-10 PK-CT-2000-HUV-50


0.1 ml 0.5 ml

PK-CT-2000-HEP-10 PK-CT-2000-HEP-50


0.1 ml 0.5 ml

PK-CT-2000-MAC-10 PK-CT-2000-MAC-50

0.15 ml 0.75 ml 5 x 0.75 ml

PK-CT-2000-NEU-015 PK-CT-2000-NEU-075 PK-CT-2000-NEU-375

0.5 ml 1 ml

PK-CT-2000-InC-50 PK-CT-2000-InC-100

0.75 ml 1.5 ml

PK-CT-2000-CL-075 PK-CT-2000-CL-150

0.2 ml 0.75 ml 5 x 0.75 ml

PK-CT-2000-RNA-50 PK-CT-2000-RNA-200 PK-CT-2000-RNA-1000

24 reactions 24 Single Use Vials 96 reactions 96 Single Use Vials

PK-CT-2000-POL-24 PK-CT-2000-POL-24Q PK-CT-2000-POL-96 PK-CT-2000-POL-96Q

PromoFectin-Neuron

PromoFectin-Insect PromoFectin-CellLine PromoFectin-siRNA


9 Cell Transfection

161

9.2 Magnet Assisted Transfection (MATra) Magnet Assisted Transfection

Transfection of adherent and suspension cells including primary cells Suited for plasmids, oligonucleotides, siRNA, and adenovirus/retrovirus transfection For stable and transient transfections Also suited for high-thoughput transfection

Adherent Cells

Suspension Cells

Magnet Assisted Lipofection Adherent Cells

Use MATra-S Immobilizer to make the cells adherent.

Suspension Cells Use MATra-S Immobilizer to make the cells adherent.

MATra-A or MATra-si Reagent

MA Lipofection Enhancer and lipofection reagent of choice

Transfected Cells

Transfected Cells

Features

Easy, fast, and reproducible procedure Functional with and without serum and antibiotics Very high transfection rates also with many hard-to-transfect cell types Up to several thousand fold increased transgene expression Very low amounts of nucleic acid necessary for transfection

Magnet Assisted Transfection (MATra) is a very fast, easy-to-handle, and highly efficient technology for transfecting cells in culture. It can be used for cells from a broad variety of species. The major components of MATra are coated magnetic nanoparticles, which effectively bind either nucleic acids or liposomes. Using a special magnet plate, the nanoparticles with the bound nucleic acids or liposomes are rapidly drawn towards the cells and are deposited at a high concentration on the outer cell membrane. Afterwards, the cells endocytose the particles, which leads to a high transfection rate even with problematic cells.

MATra nanoparticles

The MATra technology is distributed for IBA GmbH, Germany. For research use only!

Two approaches are possible: Magnet Assisted Transfection using MATra-A or MATra-siRNA Reagent. Magnet Assisted Lipofection using the MA Lipofection Enhancer and a lipofection reagent of choice. Both techniques can be used with adherent cells as well as with suspension cells. For suspension cells, the cells have to be immobilized at the bottom of the wells to make use of the magnetic field. This can be achieved by either temporarily immobilizing the cells with MATra-S Immobilizer or by using modified culture vessels. MATra is also suited for highthroughput transfection assays using adapted protocols.

DNA

DNA + nanoparticle complex

Magnet Magnet Plate

Adherent cells

Fig. 11: Magnet Assisted Transfection (MATra) principle.

MATra-A Reagent The MATra-A Reagent contains magnetic nanoparticles with an innovative polyelectrolyte coating, which effectively bind nucleic acids (e.g. plasmid DNA or oligonucleotides) by means of electrostatic interactions. Using this technique, nucleic acids are initially mixed with specifically coated magnetic nanoparticles. The magnetic particles loaded with the nucleic acids are then added to cells in culture and a strong magnetic force is applied beneath the cells. Exploiting this strong magnetic field, the full nucleic acid dose is rapidly drawn towards the target cells on the bottom of the multiwell plate or cell culture flask. This results in very close contact between the cell membrane and the highly concentrated nucleic acid loaded magnetic complexes allowing optimal nucleic acid delivery. MATra allows efficient transfection without disturbing the cell membrane architecture or leaving a hole in the cell membrane as with other transfection technologies. The transfection rate consequently is clearly increased - also with difficult-to-transfect cells such as primary cells. In addition, the amounts of nucleic acids required for successful transfection can be significantly reduced, which in turn further reduces cytotoxicity.

Cell Transfection

Applications

162

9 Cell Transfection

GFP fluorescence

Overlay

Fig. 12: Mouse fibrosarcoma cells transfected with GFP plasmid using MATra (Data kindly provided for IBA GmbH by Dr. Lutz Thon and Dr. Dieter Adam, Institute for Immunology, University Schleswig-Holstein, Kiel, Germany).

MATra-siRNA Reagent siRNAs are small interfering/inhibitory sequence-specific RNA molecules that bind to a complementary sequence of a target mRNA, block translation of that target mRNA, and initiate its degradation. Consequently, synthesis of the corresponding protein of interest is prevented and the specific gene function is knocked-out. This process is termed RNA interference or gene-silencing and it is used to elucidate gene functions.

Specifically designed small inhibitory RNA oligonucleotides (siRNAs) allow transient inhibition of gene expression in vitro and in vivo. MATra-siRNA contains specifically coated magnetic nanoparticles, which are optimized to effectively bind and stabilize siRNA. It provides optimal delivery of siRNA into many different cell types enabling efficient and prolonged genesilencing.

Fig. 13: Human endometrial stromal cells transfected with fluorescein-labeled siRNA using MATra-siRNA Reagent (Data kindly provided for IBA GmbH by Dr. Birgit Gellersen, Endokrinologikum Hamburg, Germany).

%GFP positive cells 50

40

30

20

* See page 166 for reporter plasmids.

Reagent LF

Reagent S18

Reagent FG6

10

PromoFectin-HUVEC

Fig. 14: Comparison of transfection efficiencies of MATra, PromoFectin-HUVEC, and three other common transfection reagents recommended for primary cells. Primary human dermal microvascular endothelial cells (HDMEC, see page 17) cultured in 24-well plates were transfected with a GFP reporter plasmid*. The appropriate protocols recommended by the manufacturers were used for transfection with MATra, PromoFectin, and the competing reagents. All transfections were performed in triplicate.

MATra

Cell Transfection

Phase contrast

9 Cell Transfection

The MA Lipofection Enhancer contains coated magnetic nanoparticles which bind liposomes and any polycationic transfection reagents (e.g. PromoFectin) which in turn bind nucleic acids (e.g. plasmid DNA, oligonucleotides, siRNA). Using the MA Lipofection Enhancer, cells are transfected more efficiently than with liposomal or polycationic transfection reagents alone.

MATra-S Immobilizer Suspension cells have to be made adherent first to perform Magnet Assisted

Transfection (as well as many other conventional transfection procedures). The MATra-S Immobilizer contains coated magnetic nanoparticles which bind suspension cells. Using a special magnet plate, the magnetic nanoparticles with the bound cells are rapidly drawn towards the bottom of the cell culture vessel where they are immobilized. By combining the MATra-S Immobilizer with the other MATra reagents, suspension cells can be transfected conveniently. See www.promokine.info for a listing of more than 100 cell types successfully transfected using Magnet Assisted Transfection and Magnet Assisted Lipofection.

Liposome

Cell membrane

Magnetic nanoparticles

Fig. 15: Principle of Magnet Assisted Lipofection. The MA Lipofection Enhancer can be combined with any commercially available transfection reagent based on polycations or lipids.

Response (expression level) 200

150

MATra

100

Fig. 16: Magnet Assisted Transfection and lipofection in comparison. Luciferase expression was assayed after 24 hours. With MATra reagents, low vector doses are sufficient to obtain good results.

50 2.

tion genera

fection reagents

0.05

0.1

%GFP positive cells 10 8 6 4

Reagent LF

Reagent LF + MA Lipopfection Enhancer

IBAfect

2

IBAfect + MA Lipopfection Enhancer * See page 166 for reporter plasmids.

nts

reage

1. generation lipo

Dose (µg DNA)

Fig. 17: Comparison of transfection efficiencies of two lipofection reagents with and without MA Lipofection Enhancer. Primary human dermal microvascular endothelial cells (HDMEC, see page 17) cultured in 24-well plates were transfected with a GFP reporter plasmid*. The appropriate protocols recommended by the manufacturers were used for transfection. All transfections were performed in triplicate. IBAfect liposomal transfection reagent is also available at PromoKine. Please visit www.promokine.info for more information.

ction

lipofe

0.15

Cell Transfection

MA Lipofection Enhancer

163

164

9 Cell Transfection

Magnet Plates

24 Magnet Bar Plate (8 x 12 cm): optimized for 24-well plates. 96 Magnet Bar Plate (8 x 12 cm): optimized for 96-well plates Also available: Magnet Bar Plate Sets.

Cell Transfection

To perform Magnet Assisted Transfection, specially designed neodymium magnet plates are necessary. PromoKine offers them in different sizes, optimized for different cell culture vessels.

Universal Magnet Plate (8 x 13 cm): suitable for 6-well, 12-well, 24-well, 48-well, or 96-well plates, T75 flasks, 60 mm dishes etc. Universal Magnet Plate (26 x 26 cm): suitable for 6-well, 12-well, 24-well, 48-well, or 96-well plates, T75 flasks, 60 mm dishes, 100 mm dishes etc.

Fig. 18: Universal magnet plate (8 x 13 cm) with collection of different culture plates and MATra reagents.

Fig. 19: 96 magnet bar plate (8 x 12 cm) with 96-well culture plate.


Size

Catalog Number

MATra-A Reagent

0.05 ml 0.2 ml 1 ml

PK-CT-2001-005 PK-CT-2001-020 PK-CT-2001-100

MATra-siRNA Reagent

0.05 ml 0.2 ml 1 ml

PK-CT-2021-005 PK-CT-2021-020 PK-CT-2021-100


0.05 ml 0.2 ml 1 ml

PK-CT-2003-005 PK-CT-2003-020 PK-CT-2003-100

MATra-S Immobilizer

0.05 ml 0.2 ml 1 ml

PK-CT-2002-005 PK-CT-2002-020 PK-CT-2002-100

Universal Magnet Plate, 8 x 13 cm plate Universal Magnet Plate, 26 x 26 cm plate 24 Magnet Bar Plate, 8 x 12 cm plate 96 Magnet Bar Plate, 8 x 12 cm plate

PK-CT-2011-000 PK-CT-2012-000 PK-CT-2006-000 PK-CT-2004-000

Four 24 Magnet Bar Plates Four 96 Magnet Bar Plates Two 24 and two 96 Magnet Bar Plates

PK-CT-2008-000 PK-CT-2009-000 PK-CT-2010-000

Magnet Plate

Set of four Magnet Bar Plates

Note: Please contact us to request a test offer for MATra.

9 Cell Transfection

165

9.3 Transfection Accessories

Applications Enhancement of gene expression in transfected cells

Fast and easy protocol Optimized conditions for popular cell lines Efficient for both transient and stable transfections

Features

Up to 12-fold increased gene expression levels in diverse cell lines after transfection Compatible with all commonly used cell transfection reagents

The GeneBooster Reagents are chemical cocktails designed to significantly enhance gene expression levels in transfected cells. Three unique formulations have been developed for different cell types. Simply add GeneBooster reagent

to the culture medium four hours after transfection and an increase in expression levels up to 12-fold can be achieved. The reagents can be used in combination with all commercially available DNA transfection reagents (including all PromoFectin Reagents). One vial of GeneBooster Reagent is sufficient for 150 transfections using 1 ml transfection volume. The three GeneBooster Reagents are also available in a kit.

Fig. 20: K562, HeLa-S3, MCF7, and P19 cells were transfected with the beta-Gal reporter plasmid pPK-CMV-R1 (see page 166). After 4 hours, the appropriate GeneBooster reagent was added to the cells which were stained 48 hours post transfection using a beta-Gal Staining Kit (see page 107) and examined under a microscope.

9.3.2 Cell Clean Applications Highly efficient cleanup of transfected cells

Features Efficient removal of extracellular lipid/ DNA complexes after cell transfection Non-toxic to cells · Fast and easy procedure Better downstream results post transfection CellClean is a unique washing buffer that has been designed to remove all com-

plexes of DNA and cationic lipids that associate with cell surfaces during transfection. It shows no cytotoxicity and can distinguish between extracellular and intracellular plasmid DNA. When CellClean is used as part of the cell washing procedure, more precise quantitation of the amount of DNA delivered into cells can be achieved. Moreover, it improves downstream results such as DNA stability, cytometry, and gene delivery studies. Fig. 21: Adherent cells were incubated with cationic lipid/DNA complexes for 4 hours and washed with PBS (lane 1) or CellClean (lane 2). Cells were lysed, the DNA extracted and analysed by agarose gel electrophoresis. Arrows show extracellular plasmid DNA that could be removed from cell surfaces when using Cell Clean.


Size

Catalog Number

GeneBooster Reagent I

3 vials x 1.5 ml each

PK-CT-2020-1

GeneBooster Reagent II


PK-CT-2020-2

GeneBooster Reagent III


PK-CT-2020-3

GeneBooster Reagent Kit


PK-CT-2020-0

100 ml

PK-CT-1000-01

CellClean

Cell Transfection

9.3.1 GeneBooster

166

9 Cell Transfection

Cell Transfection

9.4 Reporter, Expression and Fusion Vectors Applications

Features

Transient high-level expression of five popular reporter genes Constitutive high-level target gene ex pression yielding native or HA-tagged proteins Constitutive high-level GFP or Lucifer- ase fusion protein expression Monitoring transfection efficiency and gene expression Creating GFP or Luciferase expressing stable cell lines

Optimized CMV promoter provides high level gene expression Suited for a wide variety of cell types for both stable and transient gene expression Small vector sizes ensure high transfection efficiency and high plasmid yield in E. coli Convenient multiple cloning site with most commonly-used restriction sites for optimized cloning and expression of target genes

The pPK-CMV Vectors contain a modified human cytomegalovirus (CMV) based promoter/enhancer sequence that has been optimized for high level gene expression. Sequence motives that limit expression have been removed, while others that promote high expression levels have been retained. Consequently, these vectors yield significantly higher expression levels than other expression vectors based on the standard CMV promoter. In addition, the plasmid backbones are optimized to allow for very small vector sizes and higher plasmid yields when amplified in E. coli.

Please check our website at www.promokine.info for details and prices.

9.4.1 pPK-CMV Reporter Vectors The pPK-CMV Reporter Vectors have been designed for high level, transient expression of reporter or target genes in a wide variety of mammalian cells and tissues (in vitro or in vivo). The vectors are available without a reporter gene to clone and express a gene of interest

(blank vector) as well as with the following reporter genes: β-galactosidase (lacZ), green fluorescent protein (GFP), luciferase (Luc), chloramphenicol acetyltransferase (CAT), and secreted alkaline phosphatase (SEAP).

See also page 107/108 for luciferase, beta-galactosidase and GFP reporter assays.

Plasmid maps, multiple cloning sites, and DNA sequences are available on our website.

9.4.2 pPK-CMV Expression Vectors The pPK-CMV Expression vectors have been designed for constitutive high-level transgene expression. Three versions of the pPK-CMV Expression Vector are available: pPK-CMV-E1 does not have an expression tag. It is intended for expressing recombinant proteins in the

native forms. pPK-CMV-E2 comes with an N-terminal HA (hemagglutinin) fusion tag. pPK-CMV-E3 comes with a C-terminal HA-fusion tag. The HA fusion tags allow easy detection and purification of proteins.

All three Expression Vectors come with a positive control vector containing a CAT gene.

multiple cloning sites upstream or downstream of the reporter gene. Four versions of the fusion vector are available for convenient and efficient gene cloning and expression of the target gene fused with a reporter gene: pPK-CMV-F1 (C-GFP) has a GFP coding sequence with a stop codon inframe in the MCS. The gene of interest can therefore be expressed with GFP at the C-terminus. pPK-CMV-F2 (N-GFP) has a GFP coding sequence without a stop codon inframe in the MCS. The gene of interest can therefore be expressed with GFP at the N-terminus. pPK-CMVF3 (C-Luc) has a Luciferase coding se-

quence with a stop codon inframe in the MCS. The gene of interest can therefore be expressed with Luciferase at the Cterminus. pPK-CMV-F4 (N-Luc) has a Luciferase coding sequence without a stop codon inframe in the MCS. The gene of interest can therefore be expressed with Luciferase at the N-terminus.

9.4.3 pPK-CMV Fusion Vectors The pPK-CMV Fusion Vectors have been designed for constitutive high-level expression of GFP or Luciferase fusion proteins and efficient selection in both E. coli (using kanamycin) and mammalian cells (using neomycin). An SV40 polyadenylation signal provides efficient transcription termination and polyadenylation - thus ensuring stable mRNA. A pUC origin of replication allows for high copy numbers in E. coli. The vectors can be used for either transient transfection or for generating stable cell lines expressing only the reporter gene or the fusion target-reporter gene construct. The vectors contain two

Please check our website at www.promokine.info for details and prices.

See also page 107/108 for luciferase and GFP reporter assays.

Antibodies and ELISAs Antibodies have a broad range of applications like the detection of cellular structures as well as the specific identification, localization, and purification of intracellular and extracellular biomolecules such as proteins. They are also essential tools in describing cellular processes and for diagnostic purposes such as ELISAs. Consequently, they are important for many research disciplines like apoptosis, signal transduction, neurobiology, virology, and tumor biology.

10


3

10

Antibodies and ELISAs 10.1 Mono- and Polyclonal Antibodies . . . . . . . 169 10.2 Antigens and Cell Lysates . . . . . . . . . . . . . 172 10.3 Labeled Antibodies and Labeling Kits . . . . 173 10.4 ELISA Kits . . . . . . . . . . . . . . . . . . . . . . . . . 175

10 Antibodies and ELISAs

169

10.1 Mono- and Polyclonal Antibodies ies, immunofluorescence microscopy, immunohistochemistry, immunoprecipitation or Westem blotting. The antibodies have been produced in several different hosts (e.g. rabbit, mouse, rat, goat) and show specific reactivities against antigens from different organisms (e.g. human, mouse, rat). For many of our antibodies, blocking peptides as well as cell and tissue lysates are available that can be used for functional studies or as positive controls (see page 172). Other

primary antibodies are also available as conjugates with e.g. FITC or biotin. PromoKine lists several thousand primary antibodies on its website. Please visit www.promokine.info/antibodies to find your specific antibody of interest using our convenient search function. PromoKine provides a variety of primary antibodies for many different research topics:


n Cell Signaling: Cytokines, Growth Factors, CD Antigens, soluble Receptors, and Ligands Numerous antibodies against Interleukins (ILs), Interferons (IFNs), Fibroblast Growth Factors (FGFs), Monocyte Chemotactic Proteins (MCPs), Macrophage Inflammatory Proteins (MIPs), Transforming Growth Factors (TGFs), Tumor Necrosis Factors (TNFs), Vascular Endothelial Growth Factors and

Receptors (VEGFs, VEGFRs), InsulinLike Growth Factors and Binding Proteins (IGFs, IGF-BPs), Platelet-Derived Growth Factors (PDGFs), CC Chemokine Receptors (CCRs), Bone Morphogenetic Proteins (BMPs), Matrix Metalloproteinases (MMPs), Neurotrophins (NTs), Stromal Cell-Derived Factors (SDFs),

beta-Defensins (BDs), Eotaxins, diverse CD antigens, WNTs as well as Adiponectin, BAFF, EGF, G-CSF, GM-CSF, M-CSF, HGF, EPO, Insulin, Leptin, PLGF, Resistin, RANKL, RANTES, TIMP, TPO and many more targets.

Please see www.promokine.info/antibodies/cell-signaling n Signal Transduction Numerous antibodies against Akt-1, ATF-6, BAP29 & BAP36, CIKS, CTRP, DARP, GADPH, Hrk, IKAP, IKKs, IRAKs, JNK1, MADD, MAPKAP1, MyD88,

Nanos, PAK, PTEN, Raptor, SATB1 & SATB2, SHP2, Ski, Slug, SOCS1, STAT1, Sumo, TOR, TRAF, TRPC, TSC, Wnt-10, XBP-1, XEDAR and many more targets.

Please see www.promokine.info/antibodies/signal-transduction n Apoptosis Numerous antibodies against Caspases, Calpains, Death Receptors (DRs), HDACs and HATs as well as Acinus, AIF, Annexin V, Apaf-1, APO-1/Fas, ASK1,

Bad, Bax, Bcl-2, Bid, CAD, CARD, CIDE, Cytochrome c, Daxx, DcR, DFF, DRAK, DRAM, Fas Ligand, FLASH, FLIP, ICAD, ILPIP, LAMP, NALP, NOD2, PDCD,

Please see www.promokine.info/antibodies/apoptosis n p53 Network Antibodies e.g. against p53, p53DINP1/ SIP, p53R2, PARC, PERP, PID/MTA2, ChK2, and MTBP.

Please see www.promokine.info/antibodies/p53-network

PHAP, PIST, PUMA, RAIDD, RICK, RIP3, Smac, SODD, Survivin, TACE, TCCR, TIGAR, TRAIL, TWEAK, XAF, XIAP, ZIPK and many more targets.


PromoKine offers a wide and continuously growing range of mono- and polyclonal antibodies against a variety of interesting molecules like cytokines and growth factors as well as many other important key targets in immunology, signal transduction, apoptosis, neurobiology, virology, tumor biology, autophagy, homeostasis etc. All antibodies are purified and highly active and can be used for diverse applications such as ELISAs, flow cytometry, functional stud-

170



n Neurobiology


Numerous antibodies against APH1, APP, BACE, BRSK, CAPS, Clusterin/ ApoJ, CDNF, CRMP1, DISC1, FEZ,

GFR-alpha, Grik, KIF5, KLHL1, LGI, MANF, Neuritin, Neurotrypsin, Neurturin, Nicastrin, NK3R, NPAS3, PEN2,

SAPAP, Slitrk, Syntaphilin, TDP43, TMP21, TOCA and many other targets.

Please see www.promokine.info/antibodies/neurobiology n Stem Cell Research Numerous antibodies against Bad, betaActin, ADAM10, Adiponectin, Akt1/PKB, Bone Morphogenetic Proteins (BMPs), CD44, CD90, CD105, CXCR4, DKK, Dnmt2, Elk-1, Eotaxin, Erk2, FGF-2,

GDF-9, GM-CSF, GRO-beta, HDACs, IFN-gamma, IKK-alpha, IL-1, IL-3, IL-6, IL-8, IL-12, IL-13, Lck, LRP-5/6, MCP-1, MEK1, MIP-1, MIP-2, NFkB, Nanog, Notch 1, Oct-4, PDGFR, PPAR-gamma,

PTEN, Raf-1, RANTES, SCF, SDF-1, Smad, Sox, SSEA, Stat, Synapsin, Tau, TGF-beta, TLR, TRAF6, Tubulin, VEGF, Vimentin, Wnt and many other targets.

Please see www.promokine.info/antibodies/stem-cell-research n Cancer Research Numerous antibodies against BRCA, Cathepsins, CD26, CD44, CUEDC, EMAP-2, FAP, Galectin-3, HEC-1,

ICAM-1, IRF7, JMJD, MART-1, Matrix Metalloproteinases (MMPs), MnSOD, PARC, PAX6, Podoplanin, PTEN, OCIAD,

SIPA1L, SRC3, TIMP-1, TNF-alpha & TNF-beta, VCAM-1, VEGF and many more targets.

Please see www.promokine.info/antibodies/cancer-research n Virology Numerous antibodies against HIV and Chemokine Receptors (e.g. CCRs, CX3CR1, CXCR4, DC-SIGN, DDX-3, HIV-I and HIV-II Glycoproteins, TRIM5), Herpes (e.g. TNFRSF14/HVEM), SARS and SARS Receptors (e.g. ACE2, SARS E,

SARS M, SARS Spike, SARS-NP), Anthrax (e.g. Anthrax EF, Anthrax LF, Anthrax PA, ATR/TEM8), West Nile Virus (e.g. WNV Core, WNV Env, WNV Matrix), Influenza (e.g. Influenza-A and Influenza-B), Avian Influenza (e.g. AFHA, H5N1

Neuramidase and Hemagglutinin, H5N1 NS1/NS2), Hepatitis (e.g. HCVNS3, HDV, Hepatitis B), Epstein-Barr and Dengue Virus, (e.g. EBV, Dengue type 2) and virus-related targets (e.g. Rift Valley Fever Virus, UNG1, UNG2).

Please see www.promokine.info/antibodies/virology n Toll-Like Receptor Related Numerous antibodies against Toll-Like Receptors (TLRs, TLR1-TLR11) as well

as BTK, ECSIT, MD-1 & MD-2, RIG-1, SARM, SIGIRR, ST2, TAB1, TIRAP, TIRP,

TOLLIP, TRIAD3A, TRIF, UBC13 and many other targets.

Please see www.promokine.info/antibodies/toll-like-receptor-related n Homeostasis Numerous antibodies against ATM, beta-Actin, Tumor Endothelial Markers (TEMs), BRCC, CLIP170, CSN8, EVER,

LIMP2, LSD1, MORF4, Nephrin, op18, PTK7, RAP80, RGP1, RTP801, SCO, Slc, STEAP, TNPO3, TP1, TRIM25, VPS53

Please see www.promokine.info/antibodies/homeostasis

and many more targets.


171


n Kinases & Phosphatases IKK, JIK, JKK, Lck, LIMK, LOK, LOX-1, MAPKAPK-2, Mek, MST-1, NIK, PAK, PDK1, PKA, PKC, PTP-1B, Raf1, RIPK3,

Please see www.promokine.info/antibodies/kinases-and-phosphatases n Phospho-specific Antibodies Numerous antibodies against phosphorylated AFX, Akt, ATF2, Bad, BRCA1, c-Jun, c-Myc, Elk-1, Erk1/2, FAK, GSK-3,

IRS, JKK, Lck, MAPKAPK-2, Mek1/2, MSK1, PKC, Raf, Stat, Tau, TrkA, VASP and many other targets.

Please see www.promokine.info/antibodies/phospho-specific-antibodies n Immunology Many antibodies e.g. against ADAP, Blimp-1, Carabin, CRTH2, IFN-beta,

IL-33, MICA, ORAI, STIM2, TSLP and TSLP Receptor.

Please see www.promokine.info/antibodies/immunology n Innate Immunity Many antibodies e.g. against Cathelicidin, IPR1, LGP2, MDA5, SLPI, VISA,

Unc93b, and ZBP1.

Please see www.promokine.info/antibodies/innate-immunity n Autophagy Antibodies e.g. against Ambra1, APG5 & APG7, Apo-1/Fas, ATG, Bad, Bax,

Bcl-2, Bid, Bnip3L, DRAM, IRGM, IRS-1, LAMP, Mek1, PIST, Presenilin, and PTEN.

Please see www.promokine.info/antibodies/autophagy n Subcellular Markers Antibodies e.g. against Apaf-1, Bad, Bax, Bcl-6, Bcl-xL, Bid, CREB, DKK1, Hsp90,

Oct, Syndecan-4, Toll-like Receptors (TLRs), Tubulin, and VDAC.

Please see www.promokine.info/antibodies/subcellular-markers

ROCK-1, SEK1, SLK, Trk and many other targets.


Numerous antibodies against AAK1, Akt/PKB, ALK, AMPK, ASK1, BLK, Brk, Chk2, DNA-PK, Erk, FAK, HER2/ErbB2,

172



n Anti-Tag Antibodies


Many antibodies e.g. against FLAG-Tag, GFP, RFP, GST-Tag, HA-Tag, His-Tag, Myc-Tag, and MBP.

Please see www.promokine.info/antibodies/anti-tag-antibodies n Infectious Diseases Antibodies e.g. against Anthrax, Chlamydia LPS, or Neisseria gonorrhea.

Please see www.promokine.info/antibodies/infectious-diseases n Obesity Antibodies e.g. against ELOVL6, GPAT1, Lipe/HSL and UROP11.

Please see www.promokine.info/antibodies/obesity n Miscellaneous Antibodies Antibodies e.g. against Cox, Growth Hormone (GH), HEP-1, Immunglobu-

lins, Luciferase, Megalin, MOG, MPO, Plectin, and Syndecan.

Please see www.promokine.info/antibodies/miscellaneous-antibodies

10.2 Antigens and Cell Lysates The PromoKine peptides are perfectly suited as blocking reagent in combination with the PromoKine antibodies and as a control in e.g. immunoblotting/ Western blot, ELISA, or immunohistochemistry assays. PromoKine also provides a wide range

of ready-to-use cell and tissue lysates that can be used as controls in e.g. gel electrophoresis, Western blotting, or immunoprecipitation. Moreover, they are also useful in enzymatic activity analysis, protein-protein interaction analysis, and tissue specific expression identification.

The total tissue and cell line protein lysates are available from human, mouse, and rat origin as well as from a variety of developmental stages. Furthermore, beside lysates from normal tissue also lysates from tumor tissue and diseased tissue are available.

For a complete list of our peptides and cell lysates please see www.promokine.info/antigens-and-cell-lysates or the PromoKine Price List.


173

10.3 Labeled Antibodies and Labeling Kits https://www.promokine.info/shop

n Labeled Secondary Antibodies (HRP), alkaline phosphatase (AP), or biotin. All antibodies are purified and highly active, and they can be used for

Product

Product Description

Donkey Anti-Goat antibody (pAb), Fluorescein-coupled

diverse applications including ELISAs, immunofluorescence microscopy, immunohistochemistry, and Western blotting. Size

Catalog Number*

Donkey Anti-Goat IgG conjugated to Fluorescein (H&L)

2 mg

PK-AB718-9360

Donkey Anti-Goat antibody (pAb), Texas Red-coupled

Donkey Anti-Goat IgG conjugated to Texas Red (H&L)

2 mg

PK-AB718-9370

Goat Anti-Chicken antibody (pAb), Texas Red-coupled

Goat Anti-Chicken IgG conjugated to Texas Red (H&L)

2 mg

PK-AB718-9170

Goat Anti-Mouse antibody (pAb), AP-coupled

Goat Anti-Mouse IgG conjugated to Alkaline Phosphatase (H&L, minimum cross-reactivity)

1 mg

PK-AB718-9780

Goat Anti-Mouse antibody (pAb), Biotin-coupled

Goat Anti-Mouse IgG conjugated to Biotin (H&L, minimum cross-reactivity)

2 mg

PK-AB718-9740

Goat Anti-Mouse antibody (pAb), Fluorescein-coupled

Goat Anti-Mouse IgG conjugated to Fluorescein (H&L, minimum cross-reactivity)

2 mg

PK-AB718-9760

Goat Anti-Mouse antibody (pAb), HRP-coupled

Goat Anti-Mouse IgG conjugated to Horseradish Peroxidase (H&L, minimum cross-reactivity)

2 mg

PK-AB718-9720

Goat Anti-Mouse antibody (pAb), Texas Red-coupled

Goat Anti-Mouse IgG conjugated to Texas Red (H&L)

2 mg

PK-AB718-9770

Goat Anti-Rabbit antibody (pAb), AP-coupled

Goat Anti-Rabbit IgG conjugated to Alkaline Phosphatase (H&L, minimum cross-reactivity)

1 mg

PK-AB718-9980

Goat Anti-Rabbit antibody (pAb), Biotin-coupled

Goat Anti-Rabbit IgG conjugated to Biotin (H&L, minimum cross-reactivity)

2 mg

PK-AB718-9940

Goat Anti-Rabbit antibody (pAb), Fluorescein-coupled

Goat Anti-Rabbit IgG conjugated to Fluorescein (H&L, minimum cross-reactivity)

2 mg

PK-AB718-9960

Goat Anti-Rabbit antibody (pAb), HRP-coupled

Goat Anti-Rabbit IgG conjugated to Horseradish Peroxidase (H&L, minimum cross-reactivity)

2 mg

PK-AB718-9920

Goat Anti-Rabbit antibody (pAb), Texas Red-coupled

Goat Anti-Rabbit IgG conjugated to Texas Red (H&L)

2 mg

PK-AB718-9970

Goat Anti-Rabbit antibody (pAb), HRP-coupled

Goat Anti-Rabbit IgG conjugated to horseradish peroxidase

0.5 ml

PK-AB577-6401

Goat Anti-Rat antibody (pAb), AP-coupled

Goat Anti-Rat IgG conjugated to Alkaline Phosphatase (H&L, minimum cross-reactivity)

1 mg

PK-AB718-9580

Goat Anti-Rat antibody (pAb), HRP-coupled

Goat Anti-Rat IgG conjugated to Horseradish Peroxidase (H&L, minimum cross-reactivity)

2 mg

PK-AB718-9520

PF-XXX-Goat Anti-Human IgG

Conjugated to PromoFluor dye of choice

1 mg

PK-PFXXX-AK-H1

PF-XXX-Goat Anti-Mouse IgG


1 mg

PK-PFXXX-AK-M1

PF-XXX-Goat Anti-Rabbit IgG


1 mg

PK-PFXXX-AK-R1

Rabbit Anti-Chicken antibody (pAb), AP-coupled

Rabbit Anti-Chicken IgG conjugated to Alkaline Phosphatase (H&L)

1 mg

PK-AB718-9180

Rabbit Anti-Chicken antibody (pAb), HRP-coupled

Rabbit Anti-Chicken IgG conjugated to Horseradish Peroxidase (H&L, minimum cross-reactivity)

1.5 mg

PK-AB718-9120

Rabbit Anti-Goat antibody (pAb), AP-coupled

Rabbit Anti-Goat IgG conjugated to Alkaline Phosphatase (H&L, minimum cross-reactivity)

0.5 mg

PK-AB718-9380

Rabbit Anti-Goat antibody (pAb), HRP-coupled

Rabbit Anti-Goat IgG conjugated to Horseradish Peroxidase (H&L, minimum cross-reactivity)

1 mg

PK-AB718-9320

Note: Please see our website for an up-to-date list of available labeled antibodies. *XXX- stands for the respective PromoFluor dye (e.g. PF-350, PF-405, etc.; see page 146).


PromoKine is offering secondary antibodies which are fluorescently labeled or conjugated to horseradish peroxidase

174




Product Description

Size

Catalog Number

Rabbit Anti-Goat antibody (pAb), HRP-coupled

Rabbit Anti-Goat IgG conjugated to horseradish peroxidase

0.5 ml

PK-AB577-6403

Rabbit Anti-Mouse antibody (pAb), HRP-coupled

Rabbit Anti-Mouse IgG conjugated to horseradish peroxidase

0.5 ml

PK-AB577-6402

n Protein and Antibody Labeling Kits PromoKine also offers ready-to-use Protein and Antibody Labeling Kits. These innovative kits have been

developed for convenient and highly efficient labeling of specific antibodies or target proteins with our superior

Product

Product Description

PromoFluor-XXX-Labeling-Kit PromoFluor-XXX-Labeling-Kit

PromoFluor dyes (see page 145).

Size

Catalog Number*

Kit for Labeling of Proteins/Antibodies with a Selected PromoFluor Dye

5 reactions

PK-PF-LK-XXX-05

Kit for Labeling of Proteins/Antibodies with a Selected PromoFluor Dye

10 reactions

PK-PF-LK-XXX-10

n ECL Chemiluminescence Detection Kit High resolution and sensitivity (picogram to femtogram amounts of target) Stable hard copy results on film Long signal duration enables multiple exposures Only small amounts of antibody required for strong signals Detection possible with short exposure times (minutes) Excellent price to performance ratio

PromoKine‘s ECL Chemiluminescence Detection Kit is a convenient, non-radioactive detection system containing all the reagents required for highly sensitive chemiluminescent detection of immobilized proteins (Western blotting) or immobilized nucleic acids (Southern Blotting or Northern Blotting) conjugated with HRP (Horse Radish Peroxidase). This ready-to-use kit allows the detection of

Product

Product Description

ECL Chemiluminescence Detection Kit

Kit for Chemiluminescent Detection of HRP or HRP-labeled Biomolecules

membrane immobilized specific antigens or nucleic acid sequences directly labeled with HRP or indirectly labeled with HRPtagged antibodies/streptavidin/biotin.

Size

Catalog Number

120 ml 500 ml 1000 ml

PK-MB902-500-120 PK-MB902-500-500 PK-MB902-500-1000

Also available: Prestained RTU Protein Ladders, Fluorescent FluoProtein Gel Stain and GelQuick Protein Gel Stain/Destaining Kit (see page 123).

*XXX- stands fo the respective PromoFluor dye (PF-405, PF-415, PF-488P, PF-488LSS, PF-514LSS, PF-546, PF-555, PF-633, PF-647, PF-670, PF-750, PF-770; see page 146) Please see our website for an up-to-date list of available PromoFluor Labeling Kits.


175

10.4 ELISA Kits The ready-to-use ELISA kits are carefully validated and come with a straightforward and easy-to-use manual as well as all reagents required to perform a convenient ELISA with reliable and reproducible results. The 96-well assay plate can either be used at once or a desired number of assays can be used separately as the wells in the assay plates can be detached.

economical in medium- to high-throughput applications. The EDKs are intended for experienced ELISA users and contain all required reagents to assay your target protein/peptide in ten 96-well microtiter plates (approximately 1,000 assays). We continuously add new ELISA kits to our inventory. For an up-to-date product list please visit www.promokine.info/ELISAs

In addition, we offer ELISA Development Kits (EDKs, see page 176) which are very


n Cell Signaling: Cytokines, Growth Factors, Receptors and Ligands ELISAs for detection and quantification of human, mouse, rat, monkey, and pig: Interleukins

e.g. IL-1 alpha, IL-1 beta, IL-2, IL-4, IL-6, IL-7, IL-8, IL-12, IL-13, IL-17, IL-22, and IL-33

Growth Factors & Colony Stimulating Factors

e.g. BDNF, CTGF, EGF, FGFs (including FGF-2), G-CSF, GM-CSF, HGF, IGF-1, IGF-BPs, M-CSF, beta-NGF, PDGF-BB, TGF-beta, VEGF-A, and VEGF-C

Interferons

e.g. IFN-alpha, IFN-gamma

Chemokines

e.g. CRG-2, CTACK, Eotaxin-2, Fractalkine, GRO-alpha, GRO-beta, HCC-4, IP-10, MCP-1, MCP-2, MCP-5, MDC, MIG, NAP-2, LIX, MIP-1, MIP-3, PARC, RANTES, and SDF-1 alpha

Tumor Necrosis Factors

e.g. TNF-alpha and TNF-beta

We also supply ELISAs for Angiogenin, Angiopoietin-1, Angiostatin, BAFF, Bone Morphogenetic Proteins (BMPs), CRP, Matrix Metalloproteinases (MMPs),

EPO, flt-3 Ligand, Galectins, ICAMs, Leptin, LIF, MPO, MSP, Myostatin, PAI-1, Resistin, sRANKL, SCF, Selectins, TNF Receptors, TIMP-1 & TIMP-2, TPO,

Please see our website for a complete list of our Cell Signaling ELISAs. n CD Marker ELISAs for detection and quantification of human, mouse and monkey: CD8, CD23, CD26, CD28, sCD30, CD30L,

CD31, CD40, CD40L, CD44, CD50, CD54, CD62, CD102, CD106, CD134, CD137, CD144, CD154, and CD162.

Please see our website for a complete list of our CD ELISAs. n Hormones ELISAs for detection and quantification of human Relaxin, Thymosin alpha-1, Thymosin beta-4, and Thymulin.

Please see our website for an up-to-date list of our Hormone ELISAs.

VCAM-1, VEGF Receptors and many more interesting targets.


PromoKine offers a range of highly sensitive and specific ELISA kits that meet optimal quality standards. They can be used for accurate and consistent detection and quantitation of numerous interesting biomolecules in cell biology research and are applicable for different sample material (e.g. cell culture supernatant, serum and plasma) from various mammalian organisms (human, mouse, rat, monkey or pig). Some ELISAs can also be used with cell/tissue lysates, urine or stool samples.

176



n Apoptosis


ELISAs for detection and quantification of human and monkey: Annexin V, APO-1/Fas, APRIL, BAFF,

Bcl-2, Caspase-8 & Caspase-9, Cathepsin K & Cathepsin L, Cytochrome c, DcR3, Granzyme A & Granzyme B, LIGHT,

OPG, OX40, TRAIL, VE-Cadherin and many more interesting targets.

Please see our website for a complete list of our Apoptosis ELISAs. n Other ELISAs Here you will find a range of ELISAs for target molecules which play important roles in oxidative cell stress and inflammation as well as different metabolism patways and diseases in the skeletal, cardiovascular and renal systems. Other ELISAs included herein can be used for detecting and quantifying targets which

are essential markers e.g. in gastroenterology, fertility, diabetes, oncology, clinical immunology and orthomolecular medicine: ADMA, Adiponectin, Calprotectin, CRP, Myostatin, NT-proCNP, NT-proANP, Leptin, Matrix Metalloproteinases (MMPs), Osteonectin, Osteoprotegerin, Oxidized LDL, PAI-1,

sRANKL, RANTES, Thyreoglobulin, Urotensin II, VDBP, Vitamin D, Zonulin and many more interesting targets.

Please see our website at www.promokine.info/ELISAs for a complete list. n ELISA Development Kits Each of PromoKine’s ELISA Development Kits (EDKs) contains the following components to detect and quantify the target molecule in at least 10 x 96 ELISA plate wells: Capture antibody,

(biotinylated) detection antibody, avidinperoxidase conjugate and a thoroughly calibrated standard. A huge variety of natural and/or recombinant proteins that play a crucial role in cell signaling can be

assayed in a sandwich ELISA format. The EDKs come with a straightforward and easy-to-use instruction manual.

ELISA Development Kits for detection and quantification of human, mouse, and rat: Interleukins

e.g. IL-1 alpha, IL-2, IL-4, IL-6, IL-7, IL-8, IL-12, IL-13, IL-17, IL-22, IL-33


e.g. CTGF, EGF, FGF-2, G-CSF, GM-CSF, beta-NGF, PDGF-BB, PLGF-1, VEGF

Chemokines

e.g. Eotaxin, IP-10, MCP-1, MCP-2, GRO-alpha, GRO-beta, MIP-1, RANTES, SDF-1 alpha

We also supply EDKs for BD-1, BD-2, BD-3, BMP-2, CNTF, IFN-gamma, IGF-BP1, Leptin, NOV, PAI-1, Resistin,

RANKL, SCF, TNF-alpha, TPO and many more interesting targets.

Please see our website for a complete and up-to-date list of our ELISA Development Kits.

Cytokines and Growth Factors Cytokines and chemokines are small proteins secreted by a variety of cell types. They are important signal messengers mediating cell communication and activating other target cells through binding to specific receptors. Cytokines play a central role in the immune system where they induce inflammatory and immune responses or function as chemoattractants (chemokines). They also coordinate and regulate other biological processes such as cell growth and development or tissue repair.

11


3

11

Cytokines and Growth Factors 11.1 Cytokines, Growth Factors, Hormones and soluble Receptors . . . . . . . . . . . . . . . . 179 11.2 Animal-free Cytokines and Growth Factors . . . . . . . . . . . . . . . . . . . . 180 11.3 Cytokines and Growth Factors for Stem Cell Research . . . . . . . . . . . . . . . 181

11 Cytokines and Growth Factors

179

11.1 Cytokines, Growth Factors, Hormones, and soluble Receptors

Cell-based bioassays Functional studies Human, mouse, and rat cell culture supplements Immunoassays

Features High purity and biological activity Increased stability Produced in a range of different hosts Economic bulk sizes are available on request PromKine offers an exceptional variety of recombinant proteins or peptides of human, mouse or rat origin (>800 items!) which function as signaling molecules in intercellular communication and play important roles in both health and disease states. They can initiate, regulate or otherwise affect different effector mecha-

nisms within the responding target cells by binding to specific cell surface receptors and thus activating intracellular signal transduction cascades. Other cytokines mediate chemoattraction between cells (chemokines) and yet other signaling proteins included in our wide product range are essential growth factors and hormones which stimulate and regulate diverse cellular processes such as cell growth, proliferation, maturation, and differentiation. All cytokines, chemokines, growth factors, CD antigens, hormones, and soluble receptors available from PromoKine meet the highest quality standards. They are highly purified and biologically active and were produced in E. coli, yeast, insect cells, plants, or mammalian cell lines (to provide native posttranslational modifications if required). Purity is generally assessed by SDS-PAGE and HPLC analysis and is in most cases higher than 95%. Most of the prod-

ucts are delivered lyophilized (and are very stable in that format), while some come ready-to-use in a buffer solution. Reconstitution of the lyophilized proteins should be carried out in water or common buffer solutions. Products available at PromoKine include human, mouse and rat Interleukins, Colony Stimulating Factors (CSFs), Growth Factors, Interferons, Chemokines, Adipokines, Bone Morphogenetic Proteins (BMPs), Fibroblast Growth Factors (FGFs), Matrix Metalloproteinases (MMPs), CD antigens, Defensins, Tumor Necrosis Factors (TNFs), soluble receptors, hormones, extracellular binding proteins, non-inhibitory Serpins and other immunoregulatory proteins. We continuously add new recombinant proteins to our inventory. For an up-to-date product list please visit www.cytokines.info

https://www.promokine.info/shop Interleukins

e.g. IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-15, IL-16, IL-17, IL-19, IL-20, IL-21, IL-22, IL-24, IL-31, IL-32, and IL-33

Growth and Differentiation Factors

e.g. BDNF, CTGF, CTGFL, EGF, EG-VEGF, FGFs (including FGF-1, FGF-2 and FGF-4), GDFs, GDNF, HB-EGF, HDGF, HGF, IGF-1, IGF-2, IGF-BPs, KGF, beta-NGF, PDGFs (including PDGF-AA and PDGF-BB), PLGFs, TGF-betas (including TGF-beta 1), and VEGFs (including VEGF-A)

Colony Stimulating Factors

e.g. G-CSF, GM-CSF, and M-CSF

Interferons

e.g. IFN-alpha, IFN-beta and IFN-gamma

Chemokines

e.g. BCA-1, BRAK, CTACK, CXCL16, ENA-78, Fractalkine, GCP-2, Eotaxin, Exodus, GROalpha, GRO-beta, GRO-gamma, HCC-1, IP-10, I-TAC, LEC, LIX, Lymphotactin, MCP-1, MCP-2, MCP-3, MCP-4, MCP-5, MDC, MEC, MIG, MIP-1, MIP-3, MIP-4, NAP-2, PF-4, RANTES, SDF-1 alpha, SDF-1 beta, TAFA, TARC, and TECK

Tumor Necrosis Factors

e.g. TNF-alpha and TNF-beta

Bone Morphogenetic Proteins

e.g. BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, and BMP-7

Matrix Metalloproteinases

e.g. MMP-1, MMP-2, MMP-3, MMP-7, MMP-8, MMP-9, and MMP-13

Hormones

e.g. Adiponectin, CNTF, EPO-alpha, FSH, GH, Ghrelin, Insulin, Leptin, Prolactin, Relaxin, Resistin, and TPO

CD Antigens

e.g. CD1A, CD2, CD4, CD9, CD11b, CD14, CD22, CD23, CD25, CD29, CD31, CD34, CD43, CD45, CD54, CD56, CD62, CD68, CD72, CD95, CD99, CD100, CD105, CD106, CD120, CD124, CD137, CD141, CD146, CD147, CD152, CD154/CD40L, CD269, and CD283

We also supply other important signaling proteins/peptides such as: Activin A, APRIL, Angiogenin, Angiopoietin-1, Angiostatin, Apolipoproteins, Artemin, BAFF, Betacellulin, Cardiotrophin-1, CRP, DKK-1, Endostatin, FGF

Receptors, flt-3 Ligand, Follistatin, Galectins, Granzyme B, Heregulin, ICAMs, LIF, Maspin, Midkine, Myostatin, Nanog, Neuregulin, Neuroglobin, Neuroserpin, Neurotrophins, NOGGIN, Oncostatin M, OPG, Osteonectin, Osteopontin, PAI-1

Please see www.cytokines.info for a complete and up-to-date list.

& PAI-2, RANKL, RELM-beta, SCF, Selectins, TNF Receptors, TIE-1 & TIE-2, TIMP-1 & TIMP-2, TRAIL, VCAM-1, VEGF Receptors, Vaspin, Vitronectin, Visfatin, Wnt-3a and many more interesting targets.


Applications

180


11.2 Animal-free Cytokines and Growth Factors Applications Cell-based bioassays Functional studies Human, mouse, and rat cell culture supplements Immunoassays


Features High purity and biological activity Increased stability Free of any impurities derived from animals or bacteria Economic bulk sizes are available on request PromoKine’s animal-free cytokines and growth factors have been produced by molecular farming using tobacco plants

or barley grain as the production hosts which have the G.R.A.S. (Generally Recognized As Safe) status from the FDA. These cytokines are free of animal components, serum, endotoxins, and antibiotics, as well as human or animal infectious agents or other endogenous mammalian contaminants (e.g. traces of unwanted growth factors and cytokines). As these proteins have been expressed in plants, they show very low proteolytic, pyrogenic, and inflammatory activity compared to other commercially available proteins such as those produced in bacteria, yeast, insect, or mammalian cells. In addition, they retain proper eukaryotic folding and glycosylation resulting in an increased stability and biological activity. Degradation due to protease carryover as well as un-

wanted cell responses (e.g. proliferation or inflammatory cascades) or cell growth inhibition caused by contaminating serum components (serum carryover), endotoxins or bacterial, viral, and prion infectious agents are also eliminated or at least extremely reduced. PromoKine’s range of plant-derived proteins were produced under strict animal-free conditions. While avoiding any potential variables and interfering effects associated with the use of animal-derived manufacturing components they still maintain high biological activity and purity compared to the corresponding proteins produced using standard techniques. Thus, they are ideally suited for some very sensitive applications, e.g. in stem cell research.

https://www.promokine.info/shop Our range of animal-free cytokines and growth factors is continuously growing and includes: Interleukins

e.g. IL-2, IL-3, IL-4, IL-6, IL-12


e.g. FGF-2, KGF, TGF-beta 2 and TGF-beta 3, and VEGF-165

Colony Stimulating Factors

e.g. GM-CSF, and M-CSF

Interferons

e.g. TNF-alpha

Bone Morphogenetic Proteins

e.g. BMP-2 and BMP-7

We also supply other important signaling proteins/peptides such as: Activin A, Activin B, BAFF, flt-3 Ligand, Follistatin,

Growth Hormone, LIF, Myostatin, sRANKL, SCF and many more interesting targets that will come soon.

Please see www.promokine.info/AF for a complete and up-to-date list.

OD (490 nm) 1.6 1.5

Control fraction PromoKine´s animal-free VEGF-121

1.4 1.3 1.2 0.1

Fig. 1: Cell proliferation assay using PromoKine´s animal-free VEGF-121 and HUVEC cells (see page 13)

1.0 0.9 0.8 0.7 0.0

10-³

10-²

10-¹

1.0

10¹

ng/ml VEGF-121

10²

10³


181

Stem cells play a central role in the development of multicellular organisms and diverse regeneration processes throughout their lifes, e.g. replacing diseased and damaged tissues. They are unspecialized precursor cells characterized by their abilities of self-renewal and pluripotency, that means they can undergo numerous rounds of cell division maintaining the undifferentiated state and are able to develop into different types of cells such as skin, muscle or blood cells.

Mammalian stem cells are generally comprised of two types: Embryonic stem cells (ESCs) are derived from the epiblast tissue of the inner cell mass of an early stage embryo (blastocyst) and are considered pluripotent and capable to differentiate into any cell type of the adult body. Adult stem cells (also known as somatic stem cells) are undifferentiated cells found in the tissues of children and adults. Their main function is to replace dead or damaged specialized cells and to maintain the normal turnover of re-

generative organs. Most adult stem cells are lineage-restricted (multipotent) and characterized by a limited self-renewal and differentiation capacity usually restricted to cell types of the tissue in which they reside. In contrast, induced pluripotent stem cells (iPSCs) are not adult stem cells but specialized somatic adult cells (e.g. epithelial cells) that have been genetically reprogrammed to pluripotent cells which are in many respects similar to natural pluripotent stem cells such as embryonic stem cells.

https://www.promokine.info/shop PromoKine provides a wide range of cytokines and growth factors (>300) which are important in stem cell research: Mesenchymal Stem Cell Growth and Differentiation Factors

e.g. EG-VEGF, FGF-4, flt-3 Ligand, GDFs, GM-CSF, HGF, IFN-gamma, IL-6, SDF-1 alpha, TGF-beta 2, and TGF-beta 3

Cytokines & Growth Factors for General Propagation and Differentiation of Various Adult Stem Cell Types

e.g. BAFF, BDNF, beta-NGF, BMP-2, BMP-4, BMP-7, EGF, FGF-2, flt-3 Ligand, G-CSF, GM-CSF, IGF-1, IL-3, IL-6, IL-11, M-CSF, PDGF-AA, PDGF-BB, SCF, TARC, TGF-beta 1, TPO, and VEGFs (e.g. VEGF-A and VEGF-C)

Cytokines for Propagation of Undifferentiated Pluripotent Human Embryonic Stem Cells

e.g. Ang-1, LIF, Nanog, Wnt-1, and Wnt-3a

Hematopoietic Stem Cell Research

e.g. Ang-1, EPO-alpha, flt-3 Ligand, G-CSF, GM-CSF, Granzyme B, IFN-gamma, IGF-1, IL-2, IL-3, IL-4, IL-6, IL-7, IL-8, IL-10, IL-12, PDGF-AA, PDGF-BB, SCF, SDF-1 alpha, TGF-alpha, TGF-betas (1-3), TNF-alpha, TPO, and VEGFs (e.g. VEGF-121 and VEGF165)

General Embryonic Stem Cell Research

e.g. Activin A, Activin B, Ang-1, BDNF, BMP-4, BMP-7, EGF, FGFs (e.g. FGF-2 and FGF-4), GDF-3, Heregulin-beta, IGF-1, KGF, Noggin, Oncostatin M, TGF-alpha, TGF-beta 2, TGFbeta 3, and Vitronectin

Neural Stem Cell Research

e.g. FGF-2, EGF, Galectin-1, and Wnt-3a

Please see www.promokine.info/stemcells for a complete and up-to-date list. Note: Find more products for stem cell research (Antibodies, Stem Cell Fate Regulators, etc.) on our website.


11.3 Cytokines and Growth Factors for Stem Cell Research

Microbial Detection and Elimination Cell culture contamination is a common problem that impacts many biochemical and immunological properties of cells and it leads to unsatisfactory and unreliable results. Therefore, regular testing of cell cultures for mycoplasmas/bacteria contamination is essential. PromoKine provides highly sensitive and user-friendly test kits for this purpose, as well as products for the effective elimination and prevention of mycoplasma and eubacteria contamination.

12


12

Microbial Detection and Elimination 12.1 Mycoplasma Test Kits . . . . . . . . . . . . . . . 185 12.2 Mycoplasma Elimination Solutions . . . . . . 186 12.2.1 Mycoplasma-Ex . . . . . . . . . . . . . . . . . . . . . 187 12.2.2 BIOMYC Antibiotic Solutions . . . . . . . . . . . 187

12.3 Bacteria Test Kit . . . . . . . . . . . . . . . . . . . . 188 12.4 Disinfectants . . . . . . . . . . . . . . . . . . . . . . . 189 12.4.1 Mycoplasma-Ex Spray . . . . . . . . . . . . . . . . 189 12.4.2 Pharmacidal Spray . . . . . . . . . . . . . . . . . . . 189 12.4.3 Aquaguard . . . . . . . . . . . . . . . . . . . . . . . . . 190 12.4.4 Mycoplasma-EX Wipes . . . . . . . . . . . . . . . . 190

12 Microbial Detection and Elimination

185

12.1 Mycoplasma Test Kits

Mycoplasma

detection in cell culture, virus stocks, and medium Conventional and real-time PCR

Features Highly

sensitive (detects 10 - 15 mycoplasmas per sample volume) Fast, reproducible, and reliable results Applicable for all cell/tissue culture relevant mycoplasma species Positive and internal controls included Convenient protocol and easy sample preparation Mycoplasmas are parasitic bacteria that frequently contaminate tissue cultures and consequently also virus stocks, vaccines, and other biological materials produced in cells. The contamination can originate from serum, laboratory staff, other contaminated cultures, or the donors from which the cells have been harvested, and it often remains unnoticed. Although mycoplasmas do not cause visible cell damage, they seriously affect cell growth and metabolism, transfection efficiency, protein and monoclonal antibody synthesis, cytokine secretion, immunological properties, signal transduction, virus proliferation and can even cause DNA and RNA damage. Since mycoplasmas are very small and lack cell walls, detection using a conventional light microscope is nearly impossible. In addition, cloudiness in the

medium, typical for bacterial or yeast contaminations, does not occur even at high mycoplasma concentrations. PCR on the other hand is a highly sensitive, specific, and rapid method for the detection of mycoplasma contamination in cell culture supernatant, cell/virus stocks, and medium. All cell/tissue culture relevant mycoplasma species can be detected in one assay including M. fermentans, M. hyorhinis, M. arginini, M. orale, M. salivarium, M. hominis, M. pulmonis, and M. pirum. In addition, Acholeplasma laidlawii, M. synoviae, and Ureaplasma species can be detected.

PCR Mycoplasma Test Kit I/C The PCR Mycoplasma Test Kit I/C utilizes conventional PCR technology for convenient and specific detection of all known cell culture-contaminating mycoplasma species. The primer set is specific for the highly conserved 16S-rRNA coding region in the mycoplasma genome. The test can be performed in 3 hours and includes internal control DNA (for verifying a successful PCR run) as well as positive control DNA. All components required for the PCR (primers, nucleotides, internal control DNA) come optimally premixed and lyophilized in PCR tubes (except the Taq polymerase). For setting up the PCR, simply cut off the number of tubes required, rehydrate with rehydration buffer (containing the Hot-Start Taq polymerase), and add the

sample to be analyzed. Samples can be easily prepared by boiling the cell culture supernatant or other sample material.

PCR Mycoplasma Test Kit I/RT The PCR Mycoplasma Test Kit I/RT utilizes Real-Time PCR technology for convenient and reproducible detection of contaminating mycoplasmas. It shows excellent sensitivity and specificity and enables reliable, quantitative PCR results quickly (making the kit particularly useful for routine testing). The primer and probe set is specific for the highly conserved 16S-rRNA coding region in the mycoplasma genome. It contains FRET probes emitting fluorescent light at 520 nm and is compatible with most common Real-Time PCR cyclers (e.g. Roche Light Cycler® 1 and 2, ABI Prism®, Cepheid Smart Cycler®, and Eppendorf Mastercycler® ep realplex). The test includes internal control DNA (for verifying a successful PCR run) as well as positive control DNA. All components required for Real-Time PCR (primers, probes, nucleotides, and internal control DNA) come lyophilized in PCR tubes, except the Hot-Start Taq polymerase which is dissolved in reaction buffer. Two kit variants (kit A for e.g. Light Cycler, kit B for e.g. ABI Prism) are provided which have been optimized for different Real-Time PCR instruments. For recommended kit variant, please see www.promokine.info where you can download the manual.

PCR 100 µl cell culture supernatant

2 µl supernatant

Analyze 5 µl of the PCR sample

Boil for 10 minutes and centrifuge briefly

Fig. 1: Mycoplasma detection with the PromoKine PCR Mycoplasma Test Kit I/C is easy and convenient: Boil 100 µl cell culture supernatant or other sample material for 10 minutes; centrifuge briefly and add 2 µl supernatant to the reaction cup with the rehydrated PCR components and Taq polymerase; start PCR run; analyze PCR products by agarose gel electrophoresis. When using the PCR Mycoplasma Test Kit II, the sample is directly added to the ready-to-use, complete reaction mix to set up the PCR.


Applications

186


PCR Mycoplasma Test Kit II

The kit includes all components required for PCR (nucleotides, primers, a highperformance Taq polymerase, magnesium and an agarose gel loading dye) premixed at optimized concentrations in a ready-to-use master mix. The test takes approximately five hours to obtain clear results and reliably detects all


The PCR Mycoplasma Test Kit II utilizes conventional PCR technology for convenient and reliable detection of mycoplasmas. The primer set is specific for the highly conserved 16S-rRNA coding region in the mycoplasma genome.

M

1

2

3

4

5

mycoplasma species commonly found in contaminated cell cultures. The kit also contains positive control DNA for easy result verification but, different to the PCR Mycoplasma Test Kit I/C, contains no internal control DNA.

Fig. 2: Detection of mycoplasmas with PCR Mycoplasma Test Kit I/C. M: 100 bp DNA ladder 1: Positive control 2-6: 10; 100; 1,000; 10,000; 100,000 mycoplasmas/sample volume Internal control DNA: 479 bp band Positive sample/control: 270 bp band

6

479 bp 270 bp


Size

Catalog Number

PCR Mycoplasma Test Kit I/C

24 tests 48 tests 96 tests

PK-CA91-1024 PK-CA91-1048 PK-CA91-1096

PCR Mycoplasma Test Kit I/RT; Variant A

25 tests

PK-CA91-3025A

PCR Mycoplasma Test Kit I/RT; Variant B

25 tests

PK-CA91-3025B

PCR Mycoplasma Test Kit II

20 tests

PK-CA20-700-20

Mycoplasma fermentans DNA Standard (for qPCR)

100 µl

PK-CA91-0117

Mycoplasma orale DNA Standard (for qPCR)

100 µl

PK-CA91-0112

RTU DNA Ladder I (100 bp)

0.5 ml

PK-MB713-001N

RTU DNA Ladder II (1000 bp)

0.5 ml

PK-MB713-002N

Please see page 125 for the ready-to-use DNA Ladders.

12.2 Mycoplasma Elimination Solutions Mycoplasma contamination seriously affects cell growth and cell physiology leading to biased and unreliable experimental results. When mycoplasma contamination is detected, the contaminated cells are ideally discarded and all equipment, e.g. work benches, CO2 incubators, etc.,

are disinfected (see page 189 for disinfectant sprays). However, if the affected cells are unique or the result of extensive work, thorough elimination of the contaminating mycoplasmas is necessary. Standard antibiotics commonly used in cell culture (e.g. Pen/Strep) do

not affect mycoplasmas. Therefore, PromoKine offers two highly effective methods to get rid of mycoplasma contamination: Mycoplasma-EX, a combination of non-antibiotic and antibiotic reagents, and BIOMYC 1-3, three highly effective antibiotics.


187

12.2.1 Mycoplasma-EX

Mycoplasma

elimination in cell/tissue culture Mycoplasma elimination in virus stocks

Features Highly

effective against mycoplasma species commonly found in cell cultures Very short treatment time Very low cytotoxicity Ready to use Easy handling Mycoplasma-EX combines a non-antibiotic and an optimized antibiotic treat-

ment for the effective elimination of mycoplasmas and related microorganisms from cell/tissue culture and virus stocks. It eliminates mycoplasmas by directly killing them and not just by inhibiting their growth, with minimal toxicity for the cells in culture. Mycoplasma-EX is effective after a single treatment and destroys mycoplasmas rapidly and permanently. It is applicable for most cell lines (e.g. Vero, BHK21, GBK, ML, Hep2, 293, CRFK, H9, Molt4, MT-4, Jurkat), primary cells as well as virus stocks (e.g. SHV-1, BHV-1, HSV-1, VSV, SFV, FCV, MEV) and is highly effective against M. orale, M. hyorhinis, M. hominis, Acholeplasma laidlawii, M. arginini, M. fermentans, and other

mycoplasma species encountered as contaminants in cell cultures.

Fig. 3: Fluorescence microscope image of cells contaminated with mycoplasmas. Picture kindly provided by M. Ebert, Forschungszentrum für Medizintechnik und Biotechnologie, Erfurt, Germany.

12.2.2 BIOMYC Antibiotic Solutions Applications Mycoplasma

elimination in cell/

tissue culture

In addition, they frequently just reduce the concentration of mycoplasmas rather than completely eliminating them. As soon as treatment is discontinued, contamination will recur.

Features BIOMYC-1 and 2 Highly

effective against mycoplasma species commonly found in cell cultures Low cytotoxicity Ready to use Easy handling Antibiotics that impact bacterial cell walls (e.g. penicillin) are ineffective against mycoplasmas which lack a cell wall. Other antibiotics such as tylosin, neomycin, tetracycline, or gentamicin are only effective against a few mycoplasma species commonly found in cell cultures.

BIOMYC-1 and 2 are two antibiotics which eliminate mycoplasmas very efficiently when used in combination. After a four-day treatment with BIOMYC-1, a three-day treatment with BIOMYC-2 follows. This is repeated 2 to 3 times. Following this protocol, mycoplasmas will not develop resistance to these antibiotics - a common occurrence with other antibiotic treatment methods. BIOMYC-1 is based on the antibiotic tiamutin which is produced by the fungus Pleurotus mutilus. BIOMYC-2 is based

on the antibiotic minocycline, an improved derivative of tetracycline.

BIOMYC-3 BIOMYC-3 is based on the antibiotic ciprofloxacin which is a member of the fluoroquinolone group and inhibits synthesis of DNA gyrase, an enzyme necessary for bacterial DNA supercoiling. Many mycoplasma species are sensitive to BIOMYC-3, including M. orale, M. hyorhinis, M. fermentans, and M. arginini, and also Acheloplasma laidlawii. These species are responsible for most mycoplasma contaminations in cell cultures. No cytotoxic effects on the cells in culture have been found at the concentrations recommended for use, and the treatment can be performed within 12 days.


Size

Catalog Number

3 treatments

PK-CC91-4003

BIOMYC-1

10 ml 20 ml 100 ml

PK-CC03-036-1D PK-CC03-036-1C PK-CC03-036-1B

BIOMYC-2

10 ml 20 ml 100 ml


BIOMYC-3

10 ml 20 ml 100 ml


Mycoplasma-EX


Applications

188


12.3 Bacteria Test Kit Applications Bacteria detection in cell culture, virus stocks, and medium

Features Highly


sensitive (detects 12 bacteria genomes per sample volume) Reproducible and reliable results Applicable for 45 bacteria species Positive and internal controls included Convenient protocol and easy sample preparation

cobacterium, Peptostreptococcus, Pseudomonas, Porphyromonas, Prevotella, Salmonella, Serratia, Staphylococcus, Streptococcus, etc.). The kit provides a highly sensitive, easy, and rapid method to detect bacterial infection in various biological materials, including cell cultures. Detection requires as little as 12 bacteria genomes (~52 fg) per sample volume. The protocol can be completed within 3.5 hours with a hands-on time of less than 40 minutes.

decoloration of the medium. In addition, it has been demonstrated that standard antibiotics are mostly ineffective against resistant bacterial infection but have a strong impact on the metabolism, growth, and differentiation of the cells in culture.

Bacteria are frequently found as contaminants in cell cultures. However, cell cultures sometimes lack visual signs of the bacterial contamination, such as fast

Frequent screening of cell cultures for bacterial contamination is essential to ensure reliable results. Therefore, PromoKine offers a PCR-based Bacteria Test Kit that has been developed for accurate and reliable in vitro diagnosis of 45 bacteria species which are the common contaminants in cell cultures (e.g. Actinomyces, Bacillus, Enterococcus, Escherichia coli, Fusobacterium, Klebsiella, Lactobacillus, Micrococcus, My-

The kit contains all the necessary components for setting up a PCR - including a positive control DNA as well as an internal control DNA to confirm a successfully performed PCR run. The kit does not amplify eukaryotic DNA.

PCR 200 - 500 µl cell culture supernatant

Analyze 5 µl of the PCR sample

5 µl DNA sample Extract Bacterial DNA

Fig. 4: Bacteria detection with the PromoKine PCR Bacteria Test Kit is easy and convenient: Purify bacterial DNA from 200 – 500 µl of cell culture supernatant using an appropriate DNA extraction kit; add 5 µl sample DNA to the reaction cup with the premixed PCR components and Taq polymerase; start PCR run; analyze PCR products by agarose gel electrophoresis.


Size

Catalog Number

PCR Bacteria Test Kit

24 tests

PK-CA91-2024


48 tests

PK-CA91-2048

PromoKine also offers a Bacteria Live/Dead Staining Kit (see page 105) as well as fluorescent dyes for staining of bacteria (see page 107).


189

12.4 Disinfectants Disinfection of all tissue culture equipment and work surfaces in the lab has to be performed regularly to prevent microbial contamination. PromoKine of-

fers different disinfectants ideal for laboratory surfaces and apparatuses, CO2 incubators, and water baths.

For more information please see www.promokine.info/knowledge-base

Mycoplasma-EX Spray allows safe disinfection and cleaning of all laboratory surfaces and apparatuses, including work benches, incubators, cell storage boxes, and liquid nitrogen containers. It utilizes the powerful Mycoplasma-EX reagent in combination with an alcohol-based solution. Besides its strong mycoplasmacidal activity, it is very effective e.g. against Staphylococcus aureus, Streptococcus faecalis, Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa, Candida

albicans, Mycobacteria, HBV/HIV, Papova, Adeno, Polio and Vaccinia viruses. The effectiveness of Mycoplasma-EX Spray is further enhanced by its excellent detergent properties, achieving clean and contamination-free surfaces in one step.

Applications Disinfection Disinfection

of all laboratory surfaces of laboratory apparatuses

Features Kills

mycoplasmas within seconds against most microorganisms Non-corrosive and non-cancerogenic Does not leave any residue Supplied in a refillable spray bottle Effective

https://www.promokine.info/shop Product Mycoplasma-EX Spray Mycoplasma Disinfection Kit I (Consists of Mycoplasma-Ex Spray 500 ml, Aquaguard-1, and Aquaguard-2)

Size

Catalog Number

500 ml 5L

PK-CC91-5050 PK-CC91-5050-5L

1 Kit

PK-CC91-5060

12.4.2 Pharmacidal Spray Applications

Non-corrosive

and non-cancerogenic not leave any residue Supplied in a refillable spray bottle Does

Disinfection

of incubators

Features Free

of mercury, formaldehyde, phenol, and alcohol Very effective against bacteria, fungi, spores, and viruses

Pharmacidal Spray is a mercury-, formaldehyde-, phenol-, and alcohol-free disinfectant specifically designed for the disinfection of incubators. Spraying the incubators thoroughly every two weeks is recommended.

https://www.promokine.info/shop Product Pharmacidal Spray Mycoplasma Disinfection Kit II (Consists of Pharmacidal Spray 1 L, Aquaguard-1, and Aquaguard-2)

Size

Catalog Number

1000 ml 5L

PK-CC-110100 PK-CC-110100-5L

1 Kit

PK-CC91-5070


12.4.1 Mycoplasma-EX Spray

190


12.4.3 Aquaguard Applications Disinfection

of water trays in CO2 incubators Disinfection of water baths


Features Very

effective against bacteria, fungi, spores, and viruses Non-corrosive Non-toxic and non-volatile

The water in an incubator or water bath is a major source of cell culture contamination, particularly with respect to mycoplasma contaminations, since mycoplasmas can survive in pure water. Aquaguard, a non-volatile disinfectant, is extremely effective at eliminating the microorganisms in water. In addition, it does not cause damage to the stainless steel of the incubator or any plastic interior and it does not cause any skin irritation when used as recommended.

Aquaguard-1 is intended for disinfecting water in incubators, whereas Aquaguard-2 is intended for water bath disinfection. Both solutions keep the water reliably free of undesired microorganisms.


Size

Catalog Number

100 ml 10 x 100 ml

PK-CC01-867-1B PK-CC01-867-10B

50 ml 10 x 50 ml

PK-CC01-916-1E PK-CC01-916-10E

Mycoplasma Disinfection Kit I (Consists of Mycoplasma-Ex Spray 500 ml, Aquaguard-1, and Aquaguard-2)

1 Kit

PK-CC91-5060

Mycoplasma Disinfection Kit II (Consists of Pharmacidal Spray 1 L, Aquaguard-1, and Aquaguard-2)

1 Kit

PK-CC91-5070

Aquaguard-1 (incubator) Aquaguard-2 (water bath)

12.4.4 Mycoplasma-EX Wipes Mycoplasma-EX Wipes are an effortless way to clean and disinfect a variety of laboratory surfaces such as work benches, pipettors, cell storage boxes, the incubator interior, etc. There is no need for multiple products when you have the convenience of a simple-to-use wipe. Just wipe, toss and you’re done! Similar to our Mycoplasma-EX Spray, Mycoplasma-Ex Wipes work efficiently

against mycoplasmas, fungi, bacteria, viruses and prevent any microbial contamination. Mycoplasma-Ex Wipes contain an alcohol-based, non-corrosive and non-carcinogenic solution with a reagent that kills mycoplasmas effectively and fast. Mycoplasma-EX Wipes are excellent for maintaining a clean, contamination-free work area and also allow fast and easy

cleaning and disinfection of alcoholresistant surfaces. They can be used where it is not possible to spray and where rapid drying is important. Mycoplasma-EX Wipes are supplied as single-use wipes in a convenient product safety container. Economic refill sachets are also available.

https://www.promokine.info/shop Product Mycoplasma-EX Wipes

Size

Catalog Number

120 pieces 5 x 120 pieces

PK-CC91-5055 PK-CC91-5055-5

Alphabetical Index

191

Description

Page

1-9

Description

Page

Annexin V-Biotin Detection Kit

133

Annexin V-Biotin Reagent

133

4-Bromo A-23187 free acid

118


132

5(6)-CFDA

117

Annexin V-Cy3 Detection Kit Plus

132

5(6)-CFDA, SE (CFSE)

117


133

5-(and-6)-Carboxy-2',7'-dichlorofluorescein

117


132

5-(and-6)-Carboxy-2',7'-dichlorofluorescein diacetate

117


133

5-(and-6)-Carboxyfluorescein

117

Annexin V-EGFP Detection Kit

132

5,5'-dibromo-Bapta (tetrapotassium salt)

116

Annexin V-EGFP Reagent

133

5,5'-difluoro-Bapta (tetrapotassium salt)

116

Annexin V-FITC Detection Kit

132

5,5'-difluoro-Bapta AM

116

Annexin V-FITC Detection Kit Plus

132

5,5'-dimethyl BAPTA, AM ester

116

Annexin V-FITC Reagent

133

5,5'-dimethyl BAPTA, tetrapotassium salt

116

Annexin V-PE Detection Kit

133

5-Carboxy-2',7'-dichlorofluorescein

117

Annexin V-PE Detection Kit Plus

133

5-Carboxy-2',7'-dichlorosulfonefluorescein

117

Annexin V-PE Reagent

133

5-CFDA

117

Annexin V-PE-Cy5 Detection Kit

133

5-FAM

117

Annexin V-PE-Cy5 Reagent

133

6-CFDA

117

Antibiotics and Antimycotics

97

6-FAM

118

Antibodies

169

7-AAD (7-aminoactinomycin D)

109

Antigens

172

Aortic Adventitial Fibroblasts

28

Aortic Endothelial Cells

15

Aortic Smooth Muscle Cells

37

A A-23187 free acid

118

Apoptosis

128

Accutase-Solution

95

Apoptosis antibodies

169

Acridine Orange (AO) Solution

109

Apoptosis ELISAs

176

Actinomycin D

141

Apoptosis Inducer Set

141

Actinomycin D (10 mM)

141

Apoptotic Cell Isolation Kit

141

Adipocyte Media

77


129

Adipogenic Differentiation Medium

80

Apoptotic DNA Ladder Detection Kit I plus

129

ADP Fluorometric & Colorimetric Quantitation Kit

121

Apoptotic DNA Ladder Detection Kit II

129

ADVASEP-7 (sulfobutyl b-cyclodextrin)

112

Apoptotic/Necrotic Cells Detection Kit

140

Airway Epithelial Cell Media

65

Apoptotic/Necrotic/Healthy Cells Detection Kit

140

AK Cytotoxicity Kit

106

Aquaguard-1 (incubator)

190

Akt Activated Cell Lysate

139

Aquaguard-2 (water bath)

190

Akt Activity Detection Kit

139

ATP Fluorometric & Colorimetric Quantitation Kit

121

Akt Inhibitor

139

Akt Negative Control Cell Lysate

139

Allophycocyanin

152

Alpha-Bungarotoxin (from Bungarus multicinctus)

112

Bacteria Live/Dead Staining Kit

105

AM1-43

112

Bapta (tetracesium salt)

116

AM1-44

112

Bapta (tetrapotassium salt)

116

AM2-10

112

Bapta (tetrasodium salt)

116

AM3-25

111, 112

Bapta AM

116

AM4-64

112

Bapta-FF (free acid)

116

AM4-66

112

Bapta-FF AM

116

Amino Acid Solutions

99

BCECF

110, 118

Amphotericin B Solution

97

BCECF AM

110, 118

Anisomycin

141

BCECF AM Solution

110, 118

Annexin V Binding Buffer

133

beta-Galactosidase Reporter Assay Kit

108

Annexin V Unlabeled Reagent

133

beta-Galactosidase Staining Kit

108

B

Appendix

Alphabetical Index

Appendix

192

Alphabetical Index

Description

Page

Description

Page

Beta-Secretase Fluorometric Assay Kit

119

Cardiac Fibroblasts

28

Beta-Secretase Inhibitor I

119

Cardiac Microvascular Endothelial Cells

20

Beta-Secretase Inhibitor II

119

Cardiac Myocytes

11

Betulinic acid

141

Caspase Inhibitors

136

Biocytin (e-biotinoyl–L-lysine)

113

Caspase Screening Kit

135

Biocytin-Hydrazide

113

Caspase Substrates

136


104


137


104


135

BIOMYC-1

187


137

BIOMYC-2

187


137

BIOMYC-3

187


135

Biotin PCR Labeling Kit

149


137

Biotin-4-Fluorescein

113


137

Biotin-AM3-25

112


137

Biotin-dATP (PCR)

151


135

Biotin-dCTP (PCR)

151


137

Biotin-DHPE

111


137

Biotin-dUTP (PCR)

151


135

Biotin-Ethylenediamine, hydrobromide

113


137

Biotin-Ethylenediamine, hydrochloride

113

Caspase-3 Fluorometric Assay Kit (HTS)

137

Biotin-Rhodamine 110

113


137

Biotin-X-DHPE

111


137

Biotin-XX-a-Bungarotoxin

112


135

Bis-Fura-2, hexapotassium salt

116


137

Bladder Microvascular Endothelial Cells

20


137

Bladder Smooth Muscle Cells

40


135

Blood Cell Separation Medium

84


137

Blood Cells

48


137

Blue-fluorescent Cytoplasmic Membrane Staining Kit

111


135

Bovine Pituitary Extract

99


137

B-Phycoerythrin

152


135

Brain Microvascular Endothelial Cells

22


137

Brefeldin A

141


137

Bronchial Epithelial Cells

24


135

Bronchial Smooth Muscle Cells

40

Caspase-8 Fluorometric & Colorimetric Assay Kit

137

Buffers and Salt Solutions

98


137

Caspase-8 Fluorometric Assay Kit (HTS)

137


137


135

C CaCl2-Solution

98


137

Calcein AM

110, 114

Caspases, active

136

Calcein AM Solution

110, 114

Cathepsin B Activity Assay Kit

138


110, 114

Cathepsin B&L Inhibitor Z-Phe-Phe-FMK

138

Calcium Calibration Buffer Kit

118

Cathepsin B, human (native)

138

Calpain 1, human (native)

138

Cathepsin D Activity Assay Kit

138

Calpain 1 Inhibitor Screening Kit

138

Cathepsin D Inhibitor Screening Kit

138

Calpain Activity Assay Kit

138

Cathepsin D, human (native)

138

Calpain Inhibitor Z-LLY-FMK

138

Cathepsin G Activity Assay Kit

138

cAMP Activity Assay Kit

119

Cathepsin H Activity Assay Kit

138

Camptothecin

141

Cathepsin H, human (native)

138

Camptothecin (2 mM)

142

Cathepsin K Activity Assay Kit

138

Description

Page

Description

Page

Cathepsin L Activity Assay Kit

138

Colorimetric Cell Viability Kit III (XTT)

104

Cathepsin L, human, recombinant

138

Colorimetric Cell Viability Kit IV (MTT)

104

Cathepsin S Activity Assay Kit

138

Colorimetric DNA Fragmentation Detection Kit (IHC)

129

Cathepsin S, human (native)

138

Colorimetric Glutathione Detection Kit

131

CD Antigens, recombinant

179

Colorimetric Glutathione Reductase Activity Kit

131

CD133 Progenitor Cells

53

Colorimetric GST Assay Kit

121

CD14+ Monocytes

54

Colorimetric Nitric Oxide Assay Kit

121

CD34+ Progenitor Cells

52

Complete Cell Fractionation Kit

122

Cell Dissociation Solution ACF

96

Coronary Artery Endothelial Cells

15

Cell Fractionation

122

Coronary Artery Smooth Muscle Cells

38

Cell Lysates

172

Cryo-SFM

79

Cell Pellets

91

Cycloheximide

142

Cell Proliferation Kit I (CFSE)

104

Cycloheximide (100 mM)

142

Cell Transfection

156

Cytochrome c Apoptosis Detection Kit

130

CellClean

165

Cytokine antibodies

169

CFSE; 5(6)-CFDA, SE

110

Cytokine ELISAs

175

cGMP Activity Assay Kit

119

Cytokine Mix E for HPC Expansion

81

Chemokines antibodies

169


178

Chemokines ELISAs

175

Cytokines, recombinant

179

Chemokines, recombinant

179

Cytosol/Particulate Fractionation Kit

122

Chk2 Inhibitor

43

Cytotoxicity Kits

106

Chondrocyte Media

62

Chondrocytes

12

Chondrogenic Differentiation Medium

80

Chromogenic beta-Galactosidase Substrate Sampler Kit

108

DAPI

109

c-Jun (1-79)-GST Fusion Protein

139

DAPI Solution

109

Classical Media

85

DAPI, dilactate

109

DMEM Media

85

DASPEI

109

DMEM/Ham´s F-12 Media

85

Daunorubicin.HCl

142

Ham´s F-10 Media

85

DC Base Media

82

Ham´s F-12 Media

85

DC Generation Media

82

Leibovitz L-15 Medium

85

Dendritic Cell Media

82

McCoy´s 5A Media

86

Dermal Blood Endothelial Cells

18

MEM Media

86

Dermal Fibroblasts

27

RPMI 1640 Media

86

Dermal Lymphatic Endothelial Cells

19

Cloning Vectors

166

Dermal Microvascular Endothelial Cells

17


108, 116

DetachKit 1/2

95

Coelenterazine 400a

108, 116

Detachment Reagents

95

Coelenterazine cp

108, 116

Dexamethasone

142

Coelenterazine f

108, 116

Dexamethasone (10 mM)

142

Coelenterazine fcp

108, 116

Di-12-ANEPPQ

113

Coelenterazine h

108, 117

Di-2-ANEPEQ (JPW1114)

113

Coelenterazine hcp

108, 116

Di-4-ANEPPS

113

Coelenterazine i

108, 117

Di-8-ANEPPQ

113

Coelenterazine ip

108, 116

Di-8-ANEPPS

113

Coelenterazine n

108, 117

DiA

112

Coelenterazine Sampler Kit

108, 117

DiB

112


108, 117

DiBAC4(3)

113

Colchicine

142

DiD (DiIC18[5])

111

Colorimetric Cell Viability Kit I (WST-8)

104

Dihydrorhodamine 123

109

Colorimetric Cell Viability Kit II (WST-1)

104

Dihydrorhodamine 123, dihydrochloride salt

109

+

D

193

Appendix

Alphabetical Index

Appendix

194

Alphabetical Index

Description

Page

Description

Page

DiI (or DiIC18[3])

111

ESC-Qualified Fetal Calf Serum

88

DiI in vegetable oil

111


109, 110

DiIC1(5)

113


109, 110

Dilinoleyl DiI

111

Etoposide

142

Dilinoleyl DiO

111

Etoposide (10 mM)

142

DiO (or DiOC18[3])

111

Expression Vectors

166

DiO/DPA Membrane Potential Detection Kit

113

Extracellular Matrix Proteins

99

DiOC14(3), hydroxyethanesulfonate

111

DiOC16(3)

111

DiOC2(3)

113

DiOC5(3)

113

Fetal Calf Serum

87

DiOC6(3)

109, 110, 113

FFP-18 AM

116

DiR (DiIC18[7])

111

Fibroblast Media

70

Disinfectants

189

Fibroblasts

27

D-Luciferin, Free Acid

108

Fibronectin Solutions

99

D-Luciferin, Potassium Salt

108

FIP-18 AM

116

D-Luciferin, Sodium Salt

108

Flubi-2

118

DMAO

109

Flubida-2

118

DMEM Media

85

Fluo-3 (pentaammonium salt)

115

DMNPE-caged D-Luciferin

108

Fluo-3 (pentapotassium salt)

115

DMSO, anhydrous

114, 118

Fluo-3 (pentasodium salt)

115

DNA Damage Detection Kit

121

Fluo-3 AM

115

DNA Gel Stains

125

Fluo-3 AM Solution

115

DNA Isolation

124

FluoDNA Gel Stain

125

DNA Ladder I (100 bp)

125

FluoProtein Gel Stain

123

DNA Ladder II (1000 bp)

125

Fluorescein-a-bungarotoxin

112

DNA Size Markers

125

Fluorescein-Biotin

114

Doxorubicin.HCl

142

Fluorescein-DHPE

111

DPA

112


144

DPA/Terbium for membrane fusion assay

114


104

DPX

114


104

Dulbecco’s PBS

98

Fluorometric DNA Fragmentation Detection Kit I (BrdU)

129

Fluorometric DNA Fragmentation Detection Kit II (BrdU)

129

Fluorometric DNA Fragmentation Detection Kit III (F-dUTP)

129

Fluorometric Glutathione Detection Kit

131

E

F


109, 110

Fluorometric GST Assay Kit

121

ECL Chemiluminescence Detection Kit

174

Fluorometric Nitric Oxide Assay Kit

121

ELISA Development Kits

176

Follicle Dermal Papilla Cell Media

72

ELISAs

175

Follicle Dermal Papilla Cells

30

Endothelial Cell Growth Supplement

99

Freezing Medium

79

Endothelial Cell Growth Supplement/Heparin

99

Fura-2 (pentaammonium salt)

115

Endothelial Cell Media

63

Fura-2 (pentapotassium salt)

115

Endothelial Cell Media MV

64

Fura-2 (pentasodium salt)

115

Endothelial Cells (Large Vessels)

13

Fura-2 AM

115

Endothelial Cells (Microvascular)

17

Fura-2 AM Solution

115

Endothelial Progenitor Medium

82

Fura-2FF (K Salt)

116

Epidermal Keratinocytes

32

Fura-2FF AM

116

Epidermal Melanocytes

33

Furaptra, AM ester

116

Epithelial Cell Media

65

Furaptra, tetrapotassium salt

116

Epithelial Cells

23

Furaptra, tetrasodium salt

116

+

Description

Page

G

Description

Page

HCMEC

20

HDBEC

18

G-418 Sulphate Solution

97

HDLEC

19

GelQuick Protein Gel Stain/Destaining Kit

123

HDMEC

17

Gene Cloning and Expression

166

Hematopoietic Progenitor Cell Expansion Medium DXF

81

GeneBooster Reagent I

165

Hematopoietic Progenitor Cell Media

81

GeneBooster Reagent II

165

Hepatocyte Media

73

GeneBooster Reagent III

165

Hepatocytes

31

GeneBooster Reagent Kit

165

HEPES Buffer

98

Gentamicin Solution

97

HEPES Buffered Saline Solution

98

Glutamine, L

99

HFDPC

30

Glutamine, Stable

99

HHpC

31


134

HMEpC

25


134

hMNC-CB

57


134

hMNC-PB

56


134

hMoCD14+-PB

54


134

hMo-PB

54

Green Multi-Caspase Staining Kit

134

hMSC-AT

49

Green-beta-D-Gal

108

hMSC-BM

49

Green-fluorescent Cytoplasmic Membrane Staining Kit

111

hMSC-UC

49

Growth Factor antibodies

169

HNEpC

23

Growth Factor ELISAs

175

HOB

34

Growth Factors, recombinant

179

Hoechst 33258

109, 110

Growth Supplements

99


109, 110

GSK-3a-GST Fusion Protein

139

Hoechst 33342

109, 110

GSK-3b, human, recombinant

139


109, 110

GST Inhibitor-2 (ethacrynic acid)

121

Hormone ELISAs

175

GST protein (active)

121

Hormones, recombinant

179

HPAEC

16

HPASMC

38

hPC-PL

51

H Ham´s F-10 Media

85

HPF

28

Ham´s F-12 Media

85

HPlEpC

26

Hanks’ BSS

98

HPMEC

21

HAoAF

28

HPrSMC

41

HAoEC

15

HRCEpC

26

HAoSMC

37

HREpC

25


121

HSAEpC

24

HAT protein (P/CAF, active)

121

HSaVEC

16

HBdMEC

20

HTEpC

23

HBdSMC

40

HTSMC

39

HBEpC

24

HUAEC

14

HBMEC

22

HUASMC

39

HBSMC

40

HUF

29

HCAEC

15

Human Insulin

99

HCASMC

38

Human Primary Cells

10

hCD133+-CB

53

Cardiac Myocytes

11

hCD34+-CB

52

Chondrocytes

12

HCF

28

Endothelial Cells (Large Vessels)

13

HCH

12

Endothelial Cells (Microvascular)

17

HCM

11

Epithelial Cells

23

195

Appendix

Alphabetical Index

196

Alphabetical Index

Appendix

Description

Page

Description

Page

Fibroblasts

27

Luciferase Reporter Assay Kit I (Firefly)

107

Follicle Dermal Papilla Cells

30

Luciferase Reporter Assay Kit II (Renilla)

107

Hepatocytes

31

Luciferase Reporter Assay Kit III (Firefly & Renilla)

107

Keratinocytes

32

Luciferase Reporter HTS Kit (Firefly)

107

Melanocytes

33

Lymphatic Endothelial Cells

19

Osteoblasts

34

Lymphocyte Separation Medium 1077

84

Preadipocytes

35

Skeletal Muscle Cells

36

Smooth Muscle Cells

37

M


48

M1-Macrophage Generation Medium DXF

83

Human Transferrin

99

M2-Macrophage Generation Medium DXF

83

HUtMEC

21


164

HUtSMC

41

Macrophage Base Medium DXF

83

HUVEC

13

Macrophage Detachment Solution DXF

96

HVMF

29

Macrophage Generation Media DXF

83

HWP

35

Magnet Plate

164

Hydroxystilbamidine

114

Magnetic Separator (re-usable)

141

Hygromycin B Solution

97

Mammalian Whole Cell Protein Extraction Kit

122

Mammary Epithelial Cell Media

67

Mammary Epithelial Cells

25

MATra-A Reagent

164

I, J, K, L Indo-1 (pentaammonium salt)

115

MATra-S Immobilizer

164

Indo-1 (pentapotassium salt)

115

MATra-siRNA Reagent

164

Indo-1 (pentasodium salt)

115

McCoy´s 5A Media

86

Indo-1 AM

115

Media and Sera

60

Indo-1FF (K+ Salt)

116

Media for Primary Cells

61

Indo-1FF AM

116

Chondrocyte Media

61

Insect Cell Culture Media

87

Endothelial Cell Media

63

Insulin

99

Endothelial Progenitor Medium

82

Interleukin antibodies

169

Epithelial Cell Media

65

Interleukin ELISAs

175

Fibroblast Media

70

Interleukins, recombinant

179

Follicle Dermal Papilla Cell Media

72


118, 142

Hematopoietic Progenitor Cell Media

81


118, 142

Hepatocyte Media

73

JC-1 chloride

110

Keratinocyte Media

74

JC-1 iodide

110

Melanocyte Media

75

JNK Activated Cell Lysate

139

Mesenchymal Stem Cell Differentiation Media

80

JNK Activity Detection Kit

139

Mesenchymal Stem Cell Growth Medium

80

JNK Activity Screening Kit

139

Mesenchymal Stem Cell Growth Medium DXF

80

JNK Negative Control Cell Lysate

139

Mesenchymal Stem Cell Media

80

Keratinocyte Media

74

Monocyte Attachment Medium

83

Keratinocytes

32

Mononuclear Cell Media

84


106

Myocyte Media

62

Leibovitz L-15 Medium

85

Osteoblast Media

76

L-Glutamine Solution

99

Pericyte Media

81

Live/Dead Cell Staining Kit II

105

Preadipocyte/Adipocyte Media

77

L-JNK Peptide Inhibitor I

139

Skeletal Muscle Cell Media

78

L-JNKI1 Negative Control Peptide

139

Smooth Muscle Cell Media

79

Lucifer Yellow Cadaverine

114

Melanocyte Media

75

Lucifer Yellow CH lithium salt

114

Melanocytes

33

Lucifer Yellow CH potassium salt

114

MEM Alpha Media

86

Description

Page

Description

Page

MEM Amino Acids Solution

99

Nerve Terminal Red Staining Kit II

112

MEM D-Valine, w/o L-Glutamine

86

Neuro-DiI

111

MEM Media

86

Neuro-DiO

111

MEM NEAA Solution

99

Neurogenic Differentiation Medium

80

MEM Vitamin Solution

99

NHDF

27


122

NHEK

32

Mesenchymal Stem Cell Adipogenic Differentiation Medium

80

NHEM

33

Mesenchymal Stem Cell Chondrogenic Differentiation Medium

80

Nonyl Acridine Orange (or NAO)

110

Mesenchymal Stem Cell Growth Medium

80

Nuclear/Cytosol Fractionation Kit

122

Mesenchymal Stem Cell Neurogenic Differentiation Medium

80

Nucleic Acid Isolation

124

Mesenchymal Stem Cell Osteogenic Differentiation Medium

80

Octadecyl Rhodamine B Chloride (R18)

112

Mesenchymal Stem Cells

49

Okadaic acid

142

Mesenchymal Stem Cells from Adipose Tissue

49

Orange-fluorescent Cytoplasmic Membrane Staining Kit

111

Mesenchymal Stem Cells from Bone Marrow

49

Ordering and Technical Support

205

Mesenchymal Stem Cells from Umbilical Cord Matrix

49

Osteoblast Media

76


184

Osteoblast Mineralization Medium

76

Mineralization Medium Osteoblasts

76

Osteoblasts

34


122

Osteogenic Differentiation Medium

80

Mitochondrial Apoptosis Staining Kit

130

Oxidized Glutathione (GSSG)

121

Mitochondrial DNA Isolation Kit

124

MitoGreen

110

Mitomycin C

142

MitoRed

110

PBS

98

Mitsuhashi/Maramorosh Insect Medium

87


188

Monocyte Attachment Medium

83

PCR Mycoplasma Test Kit I/C

186

Monocytes

54

PCR Mycoplasma Test Kit I/RT; Variant A

186

Mononuclear Cell Medium

84

PCR Mycoplasma Test Kit I/RT; Variant B

186

Mononuclear Cells

56

PCR Mycoplasma Test Kit III

186

Mononuclear Cells from Cord Blood

57

PCR Purification Kit

124

Mononuclear Cells from Peripheral Blood

56

Pellets

91

MQAE

117

Pen/Strep Solution

97

MSC-Qualified Fetal Calf Serum

88

Pen/Strep/Fungizone Solution

97

Mycoplasma Disinfection Kit I

189

Pericyte Media

81

Mycoplasma Disinfection Kit II

189

Pericytes

51

Mycoplasma-EX

187

Pharmacidal Spray

189

Mycoplasma-EX Spray

189

Phorbol-12-myristate 13-acetate (PMA)

142

Myocyte Media

62

Phycobiliprotein Dyes

152

Placental Epithelial Cell Media

69

Placental Epithelial Cells

26

Plasmid Vectors

166

N, O

P

NAD/NADH Quantitation Kit

121

Pluronic F-127

114, 118

NADP/NADPH Quantitation Kit

121


114, 118

Nasal Epithelial Cells

23


114, 118

NBD C6-ceramide

112

pPK-CMV-E1 Expression Vector Kit

166

NBD C6-sphingomyelin

112


166

NBD-DHPE

111


166

Nerve Terminal Green Staining Kit I

112

pPK-CMV-F1 Fusion Vector (C-GFP)

166

Nerve Terminal Green Staining Kit II

112

pPK-CMV-F2 Fusion Vector (N-GFP)

166

Nerve Terminal Green Staining Kit III

112

pPK-CMV-F3 Fusion Vector (C-Luc)

166

Nerve Terminal Green Staining Kit IV

112

pPK-CMV-F4 Fusion Vector (N-Luc)

166

Nerve Terminal Red Staining Kit I

112

pPK-CMV-R0 Reporter Vector (Blank)

166

197

Appendix

Alphabetical Index

Appendix

198

Alphabetical Index

Description

Page

Description

Page

pPK-CMV-R1 Reporter Vector (b-Gal)

166

PromoFluor-XXX-Goat Anti-Rabbit IgG

150, 173

pPK-CMV-R2 Reporter Vector (CAT)

166

PromoFluor-XXX-Labeling-Kit

148

pPK-CMV-R3 Reporter Vector (Luc)

166


109, 110

pPK-CMV-R4 Reporter Vector (GFP)

166


109, 110

pPK-CMV-R5 Reporter Vector (SEAP)

166

Prostate Smooth Muscle Cells

41

Preadipocyte Media

77

Protein and Antibody Labeling Kits

148, 174

Preadipocytes

35

Protein Extraction

122

Prestained Protein Ladder I

123

Protein Gel Stains

123

Prestained Protein Ladder II

123

Protein Size Markers

123

Primary Cells

10

Pulmonary Artery Endothelial Cells

16

PromoFectin

160

Pulmonary Artery Smooth Muscle Cells

38

PromoFectin-CellLine

160

Pulmonary Fibroblasts

28


160

Pulmonary Microvascular Endothelial Cells

21

PromoFectin-HUVEC

160

Purple-beta-D-Gal

108

PromoFectin-Insect

160


160

PromoFectin-Neuron

160


160

Rapamycin

142

PromoFectin-siRNA

160


94

PromoFluor Amino-Modified Label

147

Receptors, recombinant

179

PromoFluor Antifade Reagent

153


134

PromoFluor Carboxylic Acid

147


134

PromoFluor ChipPack

147


134

PromoFluor Maleimide

147

Red Multi-Caspase Staining Kit

134

PromoFluor NHS-Ester

147

Red-beta-D-Gal

108

PromoFluor-XXX-Goat Anti-Mouse IgG

150

Red-fluorescent Cytoplasmic Membrane Staining Kit

111


149

Reduced Glutathione (GSSG)

121


149

Renal Cortical Epithelial Cells

26


151

Renal Epithelial Cell Media

68


151

Renal Epithelial Cells

25


149

Reporter Assays

107


149

Reporter Vectors

166


151

Resazurin, sodium salt

104


151

Resistin EDK, human

105


149

RH237

113


149

RH414

113


151

RH421

113


151

RH795

113


149

Rhod-2 (triammonium salt)

115


149

Rhod-2 (tripotassium salt)

116


151

Rhod-2 (trisodium salt)

116


151

Rhod-2 AM

116


149

Rhodamine 123

110


149

Rhodamine-DHPE

111


151

Rose-beta-D-Gal

108


151

Rosiglitazone

142

PromoFluor-XXX, phalloidin conjugate

150

R-Phycoerythrin

152

PromoFluor-XXX, streptavidin conjugate

152

RPMI 1640 Media

85

PromoFluor-XXX. biotin conjugate

152

RTU DNA Ladder I

125

PromoFluor-XXX-aadUTP

151

RTU DNA Ladder II

125

PromoFluor-XXX-Goat Anti-Human IgG

150, 173

RuBP-2S

114

R

Description

Page

Description

Page

RuBP-4S

114

Thapsigargin

142

RuBPS

114

TMRE

110, 113

TMRM

110, 113

TPEN

117

Trace Element Mix

99

S Saphenous Vein Endothelial Cells

16

Tracheal Epithelial Cells

23

SCAS

112

Tracheal Smooth Muscle Cells

39

Schneider’s Drosophila Medium

87

Transfection Reagents

156

SDIP/Europium for membrane fusion assay

114

Transferrin

99

Senescence Detection Kit

106

TRITC-DHPE

111

Sera

87

Trypan Blue Solution (0.5%)

110

Set of four Magnet Bar Plates

164

Trypsin Neutralizing Solution (TNS)

96

Skeletal Muscle Cell Media

78

Trypsin Solutions

96

Skeletal Muscle Cells

36

Tyrphostin AG 1295

142

SkMC

36

Small Airway Epithelial Cell Media

66

Small Airway Epithelial Cells

24

Smooth Muscle Cell Media

79

Umbilical Artery Endothelial Cells

14

Smooth Muscle Cells

37

Umbilical Artery Smooth Muscle Cells

39

SOD Assay Kit

121

Umbilical Vein Endothelial Cells

13

Sodium bicarbonate Solution

98

Uterine Fibroblasts

29

Soluble Receptors, recombinant

179

Uterine Microvascular Endothelial Cells

21

Stable Glutamine Solution

99

Uterine Smooth Muscle Cells

41

Staurosporine

142

Villous Mesenchymal Fibroblasts

29

Stem and Blood Cells

48

Western Blotting Kits and Reagents

174

Stem Cell Research

170, 181

White Preadipocytes

35

Sulforhodamine 101

114

X-Gal

108

Sulforhodamine 101-a-bungarotoxin

112

Zinquin, ethyl ester

117

Sulforhodamine B

114

Zinquin, free acid

117

Sulforhodamine G

114

Superoxide Dismutase (SOD, human)

121

Supplements

99

SynapseGreen C1

112


112


112

SynapseGreen C3

113


113


113


113


113

SynapseRed C2M (FM5-95)

113

T Tamoxifen citrate

142

TC Water

98

Tetrabromorhodamine 123, bromide

110

Tetramethylrhodamine-a-Bungarotoxin

113

Texas Red-DHPE

111

TexasRed PCR Labeling Kit

149

TexasRed-dUTP (PCR)

151

U, V, W, X, Z

199

Appendix

Alphabetical Index

200

International Distributors

International Distributors Cat. Number Albania

Page Biomedica Shpk Zona 4, Godina 1 Kati 1, Ap. 1 1000 Tirana Albania

Cat. Number

Product Phone:

Page

Fax: Email: Internet:

+355-69 40 91 613 +355-45 00 681 [email protected] www.biomedica.co.at

Banksia Scientific Company 3/160 Lytton Road PO Box 529 Bulimba QLD 4171 Australia

Phone: Phone: Fax: Email: Internet:

1300 769944 (toll-free) +61-7-39 02 30 00 +61-7-32 17 98 69 [email protected] www.banksiascientific.com.au

Life Research Pty Ltd. 1 Dalmore Drive, Caribbean Park Scoresby, Victoria 3179 Australia

Phone: Fax: Email: Internet:

+61-3-97 64 25 66 +61-3-86 48 58 41 [email protected] www.liferesearch.com

Austria

Biomedica Medizinprodukte GmbH & Co KG Divischgasse 4 1210 Wien Austria


+43-1-29 10 75 6 +43-1-29 01 42 9 [email protected] www.biomedica.co.at

Bangladesh

KRISHGEN BioSystems Unit Nos#318/319 3rd Floor Shah & Nahar Ind. Premises Dr E Moses Road Worli Mumbai 400018 India


+91-22-66 37 29 90 +91-22-66 37 29 91 [email protected] www.krishgen.com

Bosnia-Herzegovina

Biomedica d.o.o. Zmaja od Bosne 4 71000 Sarajevo Bosnia

Mobile: Phone: Fax: Email: Internet:

+387-66 90 60 81 +387-33 26 27 25 +387-33 26 27 26 [email protected] www.biomedica.co.at

Bulgaria

Biomedica Bulgaria Ltd. Sopharma Business Towers 5 “Lachezar Stanchev” Str. Bl. B, Office 4-B6, Fl. 6 1766 Sofia Bulgaria


+359-2-44 72 833 +359-2-44 72 831 [email protected] www.biomedica-bg.eu

Canada

CEDARLANE 4410 Paletta Court Burlington, Ontario, Canada L7L 5R2 Canada


1-800-268-5058 (toll-free) +1 289-288-0020 [email protected] www.cedarlanelabs.com

MJS BioLynx Inc. 300 Laurier Blvd. Brockville, Ontario, Canada K6V 5W1 Canada


1-888-593-5969 (toll-free) +1 613 342-1341 [email protected] www.biolynx.ca

China

Jiangsu Pu Nuo Sheng (PromoCell) Biotech Ltd. Building 2, No.6 Dong Sheng Xi Road National High Tech Development Park Jiangyin, 214434 China

Phone: +86-510-81 69 51 00 Fax: +86-510-81 69 52 00 Email: [email protected] Internet: www.promocell-china.com

Croatia

Biomedica Dijagnostika d.o.o. Paljetkova 2 10000 Zagreb Croatia


+385 1 888 5727 +385 1 888 5728 [email protected] www.biomedica.co.at

Czech Republic

BIOMEDICA CS, s.r.o. Meteor Centre Office Park “B” Sokolovská 100/94 186 00 Praha 8 Czech Republic


+420 283 931 485 +420 545 214 915 +420 283 932 507 [email protected] www.biomedica.cz

Denmark

PromoCell GmbH Sickingenstr. 63/65 69126 Heidelberg Germany

Phone: +49 6221 - 649 34 0 Fax: +49 6221 - 649 34 40 Email: [email protected] Internet: www.promocell.com

Finland

Biotop Oy Tykistökatu 6B 3 20520 Turku Finland


+358-2-241 00 99 +358-2-241 01 11 [email protected] www.biotop.fi

Hungary

Biomedica Hungária Kft. Ganz u. 16 1027 Budapest Hungary


+361-22 53 85 0 +361-20 12 68 4 [email protected] www.biomedica.hu

Australia

Appendix

Product

India

DSS Takara Bio India Pvt. Ltd. A-5 Mohan Co-op Indl. Estate Mathura Road New Delhi-110044 India

Phone: Fax: Email: Email:

+91-33-30886717/21 +91-269593821 [email protected] [email protected]

KRISHGEN BioSystems Unit Nos#318/319, 3rd Floor Shah & Nahar Ind. Premises Dr E Moses Road Worli Mumbai 400018 India



Indonesia




Israel

Biological Industries Ltd. Kibbutz Beit Haemek, 25115 Israel


+972-4-996-0595 +972-4-996-0631 [email protected] www.bioind.com

Italy

VWR International PBI S.r.l. Via San Giusto, 85 20153 Milano Italy


+39-02-48 77 91 +39-02-40 09 00 10 [email protected] it.vwr.com

Japan

Funakoshi Co. Ltd. 9-7 Hongo 2-chome Bunkyo-ku Tokyo Japan 113-0033


+81-3-56 84 16 20 +81-3-56 84 17 75 [email protected] www.funakoshi.co.jp

TaKaRa Bio Inc. Seta 3-4-1 Otsu, Shiga 520-2193 Japan

Phone: Fax: Internet:

+81-77-543 7247 +81-77-543 9254 www.takara-bio.co.jp

Korea

Sungwoo Life Science Co., Ltd. Samsung BLDG 4th Floor 560-5 Uijeongbu 2Dong Uijeongbu-City Kyeonggi-Do South Korea


+82-2-985-7471 +82-31-876-9149 [email protected] www.sungwools.com

Latvia

SIA DIAMET Aizkraukles iela 23 Riga, 1006 Latvia


+371 26 47 47 64 +371 67 54 33 31 [email protected] www.diamet.lv

Macedonia

Biomedika Dijagnostika dooel Ul. llindenska br. 103 lokal 2 i 3 1000 Skopje Macedonia



Malaysia

Biomarketing Services SDN BHD 21, Jalan 4/62A Bandar Menjalara, Kepong 52200 Kuala Lumpur Malaysia


+603 6273 3068 +603 6272 0093 [email protected] www.biomarketing.com.my

Nepal




Netherlands

Bio-Connect B.V. Begonialaan 3a 6851 TE Huissen Netherlands


+3126 326 4450 +3126 326 4451 [email protected] www.bio-connect.nl

New Zealand

Banksia Scientific Company 12 Brigade Street Airport Oaks Auckland New Zealand


+64-9-921 26 26 +64-9-921 26 27 [email protected] www.banksiascientific.co.nz

Life Research Pty Ltd. Level 1, 5 Lakeside Drive Burwood East Victoria 3151 Australia


+61-3-98 03 00 45 +61-3-86 48 58 41 [email protected] www.liferesearch.com

201

Appendix


Appendix

202


Norway


Phone: +49 6221 - 649 34 0 Fax: +49 6221 - 649 34 40 Email: [email protected] Internet: www.promocell.com

Poland

Biomedica Poland sp. z.o.o. Ul. Raszyńska 13 05-500 Piaseczno Poland


+48-22 73 75 996 +48-22 73 75 994 [email protected] www.biomedica.pl

Portugal

LABCLINICS, S.A. Industria, 54 08025 Barcelona Spain


+34-93-446 47 00 +34-93-348 10 39 [email protected] www.labclinics.com

Romania

S.C.C.Y.B.E.R. Srl Romania Str. Smaranda Braescu, nr. 20D Sector 1, Cod 014203 70941 Bucharest Romania


+40-37 27 66 844 +40-37 27 66 845 +40-21 23 22 204 [email protected] www.cyber-srl.ro

Serbia

BIOMEDICA MP d.o.o. Cara Dusana 214 11080 Zemun Serbia



Singapore

Biomed Diagnostics Pte. Ltd. 18 Boon Lay Way #05-105-108 TradeHub 21 Singapore 609966


+65-6-298 43 47 +65-6-298 47 23 [email protected] http://www.biomed.com.sg/main.htm

Singlab Technologies Pte. Ltd. 1003 Bukit Merah Central #07-50 Inno. Centre Singapore 159836


+65-6-272 07 88 +65-6-270 71 28 [email protected] www.singlab.com.sg

Slovakia

Biomedica Slovakia s.r.o. Drobného 27 84101 Bratislava Slovakia


+421-2-69 30 99 01 +421-2-69 30 99 08 [email protected] www.biomedica.sk

Slovenia

Biomedis M.B. Slokanova ulica 12 2000 Maribor Slovenia


+386-2-47 16 300 +386-2-47 16 304 [email protected] www.biomedis-mb.si

South Korea

Sungwoo Life Science Co., Ltd. Samsung BLDG 4th Floor 560-5 Uijeongbu 2Dong Uijeongbu-City Kyeonggi-Do South Korea


+82-2-985-7471 +82-31-876-9149 [email protected] www.sungwools.com

Spain

LABCLINICS, S.A. Industria, 54 08025 Barcelona Spain


+34-93-446 47 00 +34-93-348 10 39 [email protected] www.labclinics.com

Sri Lanka

KRISHGEN BioSystems Unit Nos#318/319 3rd Floor Shah & Nahar Ind. Premises Dr E Moses Road Worli Mumbai 400018 India



Sweden

Mediqip AB Förrådsvägen 10 141 46 Huddinge Sweden


+46-8-448 10 10 +46-8-448 10 75 [email protected] www.mediqip.se

Taiwan

Biochiefdom International Co. Ltd. 1F, No 50, Yishou St. Wenshan District Taipei 11659 Taiwan


+886-2-29 37 00 88 +886-2-29 37 97 17 [email protected] www.bio-chief.com.tw

Thailand

Biomed Diagnostics (Thailand) Co. Ltd. 1518/4 Kengrungruengchai Tower, 7th Floor Wong Sawang, Bang Sue, Bangkok 10800 Thailand


+66-2-879 60 26 +66-2-879 60 65 [email protected] www.biomedthai.com

For an up-to-date list of all PromoCell and PromoKine distributors, please see www.promocell.com/contact

Terms and Conditions

203

General Terms

Delivery

1. These conditions of sale and delivery apply to all contracts of supply between us and our customers, even when in individual cases they are not referred to. 2. A customer’s inconsistent conditions of purchase shall have no validity, even when we do not expressly reject such conditions. 3. The customer is deemed to agree to these conditions of sale and delivery upon unconditional acceptance of the goods supplied. 4. Rights of the customer arising during the course of business with us are not transferable. 5. Any agreement that differs from these conditions of sale and delivery, or any recognition of the customer’s contradictory purchase conditions, requires our express confirmation in writing.

1. Delivery times will be specified after careful co-ordination. They are however not legally binding. Claims for compensation due to delayed or non-delivery are excluded. 2. If a delivery date specified by us is exceeded by more than two weeks, the customer is entitled to give us a further two week time limit for delivery. Should the delivery not be carried out by the end of the additional two week delivery period, the customer is entitled to rescind the contract. The rescission must be made promptly and in writing, at the latest within a week of the expiry of the additional two week delivery period. 3. All deliveries take place at the customer’s risk. The passing of risk takes place on the handing over of the goods to the carrier or haulier. The careful choice of dispatch method and means of transport is at our discretion with exclusion of any and all liability. 4. Deliveries in part are permitted. They are to be regarded as separate deliveries.

Offers 1. In the absence of a declaration to the contrary, we are not bound by any offers we make. 2. Supply contracts only take effect when confirmed by us in writing. 3. Any inquiries or complaints concerning confirmation are to be made promptly in writing. 4. Assurances and assured properties only exist when they are expressly described as such. The descriptions and other statements in our catalogues and price lists are accurate to the best of our knowledge and belief. Under no circumstances may any assurances as to properties be derived from such descriptions and statements.

Prices 1. Our prices quoted in the current price list are net prices, VAT (value added tax) not included. 2. VAT is charged according to the applicable laws in effect. 3. Charges for delivery, cooling and insurance are indicated in the valid price list. 4. PromoCell prices are subject to change without prior notice.

Payment 1. Payment is to be made without any deductions within 14 days of delivery and remittance of the bill. 2. Default of payment will result in legal proceedings upon the expiry of the payment period without further warning. In this case, or in the case of a warning, we are entitled to interest for late payment at the current normal bank borrowing rate, or alternatively at a rate of 5% above the current discount rate of the Deutsche Bundesbank. 3. A cash discount is only permitted when expressly stated on the bill and when payment is made within the stated period. 4. An offset against or demand for payment is only permitted when the customer has an undisputed or legally established counterclaim. 5. If our conditions of payment are not complied with or if justifiable doubts exist regarding the customer’s ability or willingness to pay, we are entitled to

rescind the contract and to demand the return of the delivered goods, the costs of delivery to be borne by the customer. In addition, all time outstanding claims will become due. Justifiable doubts referred to above include, but are not restricted to: unfavourable information, deterioration of financial circumstances, initiation of insolvency or bankruptcy proceedings, execution measures against the customer, protest of bills, and default of payment for any other reason. 6. If the customer does not take delivery of properly delivered goods, we are entitled, after setting an additional time limit, to rescind the contract, or to damages for non-compliance with the contract. In the latter case we are entitled to demand either damages of 25% of the sales price without proof, or the actual amount of damages.

Complaint 1. Any complaint regarding the goods, such as defects, lack of assured properties, wrong delivery and over- or underdelivery are, after immediate discontinuation of use, to be reported in writing. 2. In the case of wrong delivery, the goods will be taken back at our expense and replaced with the goods ordered. Excess deliveries may similarly be sent back to us at our expense. In the case of under delivery, the customer may choose between a reduction in payment or complete fulfilment of the order. 3. In the case of delivery of faulty goods we are entitled to replace the faulty goods with goods free of faults. Should the replacement delivery also be faulty, the customer may choose between a reduction in payment or cancellation of the contract. Claims for compensation are excluded except in the case of lack of assured properties. 4. The right to complain expires in the case of obvious or apparent defects two weeks after receipt of the goods, and in the case of non-apparent defects six months after receipt of the goods. 5. Goods may only be returned by arrangement with us and with the issue of a return number. The goods must be

Appendix


Appendix

204


properly packed. If returned goods are damaged due to the fault of the customer, the customer is obliged to pay compensation for such damage. 6. In the case of delivery to traders the customer is to inspect the goods without delay upon receipt and to report any complaints to us immediately. If the customer fails to make such a report the goods will be deemed approved. In the case of a fault that is not apparent upon inspection, the complaint must be reported immediately upon discovery of the fault, otherwise the goods will be deemed approved as regards such fault.

Use of Goods 1. Goods delivered by us are intended exclusively for the purposes of research and may not be used on human beings, animals, in the household, or for any private use. We emphasise that the use of our products for diagnostic or therapeutic purposes is subject to current legal regulations. 2. We do not accept any liability for the improper use of our products. We deliver only to commercial enterprises, resalers and public research, development and teaching establishments. Legal regulations require us to carefully check orders and to refuse delivery when there are signs of improper use of our products. Orders from students or other private individuals must be given from an official office of their institute or firm. 3. We expressly oppose research or any uses which run counter to ethical, legal or political standards or principles within Germany and/or the import country, or whose ethical justification is the subject of public debate and dispute in the respective country (e.g. certain cloning experiments). We do not accept any liability for such research or use by our customers, or by third parties who have received our products from our customers.

Liability 1. All products delivered by us are only to be handled by trained personnel who are well aware of any potential dangers. Should any liability for compensation

arise as a result of damage caused by breach of this condition, the customer shall be liable to pay any such compensation. In all cases, current safety regulations are to be observed, especially in the case of cells of human or animal origin, which always represent a potential risk of infection. The lack of a danger symbol does not mean that the substances concerned are harmless. 2. We accept no responsibility for damage, loss, or injury caused by incorrect handling or use in the household, on humans, or on animals. In as far as we have made confirmation of delivery dependent upon a specified use of certain products, the customer accepts full liability for any damage suffered by us as a result of the customer’s incorrect use of the product. In the case of products which may only be used in accordance with legal or official regulations, the customer’s order simultaneously counts as a declaration that the products will be used for a specific purpose in the above sense. 3. Our customers are required without exception to apply the laboratory guidelines of the professional association of the chemical industry when dealing with our products. 4. Passing on of our products to private individuals is not permitted. If as a result of a breach of this condition for whatever reason we should receive a claim for compensation from a private individual, we shall have a right of redress against the customer. 5. PromoCell provides no warranties as to the accuracy of the information contained in printed materials like product informations, catalogs, instruction manuals, etc. and shall bear no responsibility or liability whatsoever for any errors, faults or omissions in the information.

ers or employees. We accept no liability for damage caused by the behaviour of companies or individuals which sell our products. 3. The sale of our products under their trade names for further commercial use does not mean that these products and their trade names may be freely used in general circulation. Some products, their production and use or their trade names are patented or protected trade names. The further commercial use of such products and their trade names occurs as regards a third party under the exclusive responsibility and liability of the customer.

Indemnification

Managing Director: Dirk Hüttner AG Mannheim HRB 335480

1. In addition to section “complaint”, claims by the customer are excluded, in so far as they are not based upon intention or gross negligence. 2. In the case of delivery to a trader this exclusion also applies to any damage caused by the intention or gross negligence of our business associates, carri-

Miscellaneous 1. German law shall apply to all contracts entered into by us, including those for deliveries abroad. We expressly reject the application of the Vienna UN-agreement relating to international sales contracts of 11. 04. 80. 2. In the case of contracts with traders who do not belong to the traders specified in §4 of the code of commercial law, the place for fulfilment of contracts and the court of jurisdiction shall be Heidelberg. 3. If any of the above conditions is held to be invalid, the validity of any of the other conditions shall not be affected. 4. Only the German version of this Terms of Delivery and Payment is valid. This translation shall only serve as assistance for our customers. You can get a German version on request.


Ordering and Technical Support

205

Ordering and Technical Support Ordering Information

Phone

Fax

Online

Our customer service representatives are ready to assist you with ordering Monday through Friday

Please copy the fax order form on the last page and send it to the following fax numbers:

USA/Canada 1 – 866 – 251 – 2860 (toll free)

USA/Canada 1 – 866 – 827 – 9219 (toll free)

Online ordering is available 24 hours/day, 7 days per week. Our online ordering system is secure, fast, and convenient. Check out www.promocell.com or www. promokine.info for more information.

Germany 0800 – 776 66 23 (toll free)

Germany 0800 – 100 83 06 (toll free)

France 0800 90 93 32 (toll free)

France 0800 90 27 36 (toll free)

United Kingdom 0800 – 96 03 33 (toll free)

United Kingdom 0800 – 169 85 54 (toll free)

Switzerland +49 6221 – 649 34 0

Switzerland +49 6221 – 649 34 40

Netherlands +49 6221 – 649 34 0

Netherlands +49 6221 – 649 34 40

Email Please send an e-mail to [email protected] and include the following information: Catalog number, name, size and quantity Shipping and billing address Contact person`s name and telephone number

Other Countries Please see www.promocell.com/distributor or call +49 6221 – 649 34 0

PromoCell GmbH directly supplies to Belgium, Canada, France, Germany, Ireland, Liechtenstein, Luxembourg, Monaco, Netherlands, Switzerland, United Kingdom, and the United States. All deliveries are carried out using our recommended carrier, unless an alternative agreement has been arranged with the customer. Delivery time and shipment costs are extremely favorable. Our local sales representatives and our multilingual customer service are looking forward to serving you. For current information about PromoCell and PromoKine distributors see www.promocell.com/distributor or contact us via e-mail or phone.

Technical Support Providing excellent technical support for our products is our top priority. Our technical service team relies on years of laboratory experience to assist you with product selection and advice, to maximize product performance. You may reach our technical support via telephone/fax (see numbers on the reverse side of the catalog) or by sending an e-mail to: [email protected]

Technical support is completed by our online Knowledge Base: www.promocell.com/knowledge-base www.promokine.info/knowledge-base It provides reference literature, application notes, and a library of technical questions and answers related to our products.

Product Use Products are sold for research use only and are not intended for use in diagnostic procedures or for therapeutic and medical purposes.

Trademarks Used in this Catalog The trademarks, logos, and service marks (collectively the “Trademarks”) displayed in this catalog, including but not limited to those contained in the list below are trademarks or registered trademarks of PromoCell GmbH in Europe, the U.S. and other countries. All other brands and product names are trademarks or registered trademarks of their respective companies. Nothing contained in this catalog should be construed as granting, by implication, estoppel, or otherwise, any license or right to use any trademarks

displayed in the catalog without the permission of PromoCell GmbH or such third party that may own the trademarks displayed in the catalog. The misuse of the trademarks displayed in the catalog, or any other content in the catalog, is strictly prohibited. Please be advised that PromoCell GmbH will aggressively enforce its intellectual property rights to the fullest extent of the law, including the seeking of criminal prosecution. PromoCell™ (PromoCell GmbH) PromoFectin™ (PromoCell GmbH) PromoFluor™ (PromoCell GmbH) PromoKine™ (PromoCell GmbH) DetachKit™ (PromoCell GmbH) SupplementPack™ (PromoCell GmbH) SupplementMix™ (PromoCell GmbH) Cell Culture Assistant™ (PromoCell GmbH) ABI Prism® Alexa Fluor® Cell Lab Quanta SC® Cepheid Smart Cycler® CyTM LightCycler® Mastercycler® MitoCaptureTM Rhodamine Green® RNAlater® Texas Red®

Appendix

To place an order please use any of the following convenient options:

206

Notes

Appendix

Notes

Notes

207

Appendix

Notes

208

Notes

Appendix

Notes

Fax Order Form Shipping Address Mr.

The Cell Culture Experts www.promocell.com

Invoice Address (if different from shipping address) Ms.

Dr.

Prof.

Mr.

Ms.

First Name

First Name

Last Name

Last Name





Street, No

Street, No

ZIP, City

ZIP, City

Country

Country

Dr.

Prof.

Phone E-mail

Order Details Catalog Number

Product Name

Quantity

Your Cell Research Partner www.promokine.info

Please write in capital letters

Training Courses www.promocell-academy.com Please visit our websites regularly for upcoming new products and training courses

Please send me a copy of the

Please send me your newsletters

PromoCell catalog

PromoCell

PromoKine catalog

PromoKine

PromoCell Academy catalog

PromoCell Academy

Date, Signature

PromoCell GmbH Sickingenstr. 63/65 69126 Heidelberg Germany USA/Canada Phone: 1 – 866 – 251 – 2860 (toll free) Fax: 1 – 866 – 827 – 9219 (toll free) Deutschland Telefon: 0800 – 776 66 23 (gebührenfrei) Fax: 0800 – 100 83 06 (gebührenfrei) France Téléphone: 0800 90 93 32 (ligne verte) Téléfax: 0800 90 27 36 (ligne verte)

2013 | 2014


2013 | 2014


United Kingdom Phone: 0800 – 96 03 33 (toll free) Fax: 0800 – 169 85 54 (toll free) Switzerland Phone: +49 6221 – 649 34 0 Fax: +49 6221 – 649 34 40 Netherlands Phone: +49 6221 – 649 34 0 Fax: +49 6221 – 649 34 40 Other Countries Phone: +49 6221 – 649 34 0 Fax: +49 6221 – 649 34 40

Email: [email protected] [email protected]

www.promocell.com

Human Primary Cells & Media Stem and Blood Cells & Media Cell Biology Products