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Cytokine Immunocytochemistry Fax (65) 860-1590 Fax (49) 40 531 58 92 Fax (81) 3 541-381-55 E-Mail: [email protected]

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Cytokine Immunocytochemistry Fax (65) 860-1590 Fax (49) 40 531 58 92 Fax (81) 3 541-381-55 E-Mail: Canada@Pharmingen.Com PharMingen International United States Canada PharMingen Europe Japan PharMingen Canada Tel 619-812-8800 Asia Pacific Becton Dickinson GmbH Nippon Becton Dickinson Toll-Free 1-888-259-0187 Orders 1-800-848-6227 BD Singapore HQ PharMingen Europe Company Ltd. Tel 905-542-8028 Tech Service 1-800-825-5832 Tel (65) 860-1478 Tel (49) 40 532 84 48 0 Tel (81) 3 541-382-51 Fax 905-542-9391 Fax 619-812-8888 Cytokine Immunocytochemistry Fax (65) 860-1590 Fax (49) 40 531 58 92 Fax (81) 3 541-381-55 e-mail: [email protected] http://www.pharmingen.com Africa Germany Latin/South America Spain Becton Dickinson Worldwide Inc. Becton Dickinson GmbH BDIS (USA) Becton Dickinson S.A. 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Tel 08 007 830 373 Becton Dickinson Tel (81) 3 5256-5311 Tel (351) 1 422 0100 Technical Service (UK only) Immunocytometry Systems Fax (81) 3 5256-5370 Fax (351) 1 422 0110 Tel 08 007 830 342 Orders Tel (33) 476 68 37 32 Korea South Africa Fax (33) 476 68 35 06 Becton Dickinson Korea Inc Becton Dickinson Technical Service Tel (822) 5694030 Immunocytometry Systems Tel (33) 476 68 34 25 Fax (822) 5694048/9 Tel (27) 11 807 1531 Fax (33) 476 68 35 06 Fax (27) 11 807 1953 10975 Torreyana Rd. San Diego, CA 92121 BULK RATE U.S. POSTAGE (619) 812-8800 Tel PAID (619) 812-8888 Fax Permit No. 94 San Diego, CA 99-6081-74A Technical Assistance 1-800-TALK-TEC (825-5832) • Fax 619-812-8888 XX World Wide Web http://www.pharmingen.com Table of Contents Introduction The term cytokines refers to a broad group of signaling proteins that, in general, Introduction . 1 are produced transiently after cellular activation, act locally as autocrine, juxtacrine Immunocytochemical Measurements of Mouse Cytokines . 2 or paracrine biological response modifiers, and exert their actions by binding to specific high-affinity receptors on target cells. The vast cytokine group includes Immunocytochemical Measurements of Human Cytokines . 3 members of the interleukin, interferon, tumor necrosis factor, colony stimulating factor, transforming growth factor and chemokine families of proteins. Due to the Important Points to Consider when Choosing a critical roles played by cytokines in the regulation of immunity and inflammation, Method to Study Cytokine Production by Individual Cells . 4 there has been increased emphasis on defining the qualitative, quantitative, temporal and spatial patterns of their expression. These types of analyses have Comparison of Cytokine Immunocytochemistry and Flow benefited tremendously from the recent introduction of methods that allow for the Cytometry . .5 characterization of cytokine-producing cells at the single cell level. Such methods are often based on the use of monoclonal antibodies that are specific for a particular Relative Sensitivities of Immunocytochemistry cytokine and immunofluorescent or immunoenzymatic techniques to detect and and Flow Cytometry . 6 quantify specific antibody binding to target cytokines (Sander et al., 1991). In the latter case, immunocytochemistry (ICC) can be a powerful immunoenzymatic Critical Parameters in Immunocytochemistry . 7 technique that allows for the detection of the cytokines associated with individual cells that have been immobilized on microscopic slides. Schematic of Staining Protocol . 8-9 An important goal for PharMingen has been to develop and to provide high Analysis of Mouse TNF-α-Producing Cells quality antibodies that can be used to study cytokine expression at the single cell by Immunocytochemistry . .10 level. In this mailer, PharMingen is pleased to provide a list of selected antibody clones that are now available for the immunocytochemical staining of human Features of Intracellular Staining . 11 and mouse cytokine-producing cells. Moreover, reagents and methods are presented that are suitable for the immunoenzymatic analysis of cytokine Comparison of Immunocytochemical Detection Systems . 11 protein expression using the technique of immunocytochemistry. Cytokine Immunocytochemistry Protocol . 12-13 Product Listing . 14 Controls for Cytokine Immunocytochemistry . 15 Cytokine Immunocytochemistry Program Overview . 16 References . .16 We are thankful to Dr. Jan Andersson (Karolinska Institute, Sweden) and Dr. Thomas E. Fehniger (Astra Draco, Sweden) for consulting with PharMingen in the development of methods and reagents for the immunocytochemical analysis of cytokine-producing cells. Cover image is an artistic rendering of cytokine immunocytochemical staining and does not represent actual data. All products are for research use only. Not for use in diagnostic or therapeutic procedures. Orders 1-800-848-MABS (6227) • Phone 619-812-8800 Technical Assistance 1-800-TALK-TEC (825-5832) • Fax 619-812-8888 XXi XX1 World Wide Web http://www.pharmingen.com World Wide Web http://www.pharmingen.com Table of Contents Introduction The term cytokines refers to a broad group of signaling proteins that, in general, Introduction . 1 are produced transiently after cellular activation, act locally as autocrine, juxtacrine Immunocytochemical Measurements of Mouse Cytokines . 2 or paracrine biological response modifiers, and exert their actions by binding to specific high-affinity receptors on target cells. The vast cytokine group includes Immunocytochemical Measurements of Human Cytokines . 3 members of the interleukin, interferon, tumor necrosis factor, colony stimulating factor, transforming growth factor and chemokine families of proteins. Due to the Important Points to Consider when Choosing a critical roles played by cytokines in the regulation of immunity and inflammation, Method to Study Cytokine Production by Individual Cells . 4 there has been increased emphasis on defining the qualitative, quantitative, temporal and spatial patterns of their expression. These types of analyses have Comparison of Cytokine Immunocytochemistry and Flow benefited tremendously from the recent introduction of methods that allow for the Cytometry . .5 characterization of cytokine-producing cells at the single cell level. Such methods are often based on the use of monoclonal antibodies that are specific for a particular Relative Sensitivities of Immunocytochemistry cytokine and immunofluorescent or immunoenzymatic techniques to detect and and Flow Cytometry . 6 quantify specific antibody binding to target cytokines (Sander et al., 1991). In the latter case, immunocytochemistry (ICC) can be a powerful immunoenzymatic Critical Parameters in Immunocytochemistry . 7 technique that allows for the detection of the cytokines associated with individual cells that have been immobilized on microscopic slides. Schematic of Staining Protocol . 8-9 An important goal for PharMingen has been to develop and to provide high Analysis of Mouse TNF-α-Producing Cells quality antibodies that can be used to study cytokine expression at the single cell by Immunocytochemistry . .10 level.
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