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PDP - Template Name: Flow Cytometry Panels
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  1. Products /
  2. Flow Cytometry Kits & Reagents /
  3. Human T Cell Th1 Cytokine Response Flow Cytometry Panel

Human T Cell Th1 Cytokine Response Flow Cytometry Panel #28572

    Product Information

    Protocol

    Product Description

    The Human T Cell Th1 Cytokine Response Flow Cytometry Panel can be used to observe production of signature Th1 cytokines including IFN-γ and TNF-α.

    T cells are identified by expression of CD3. There are two major subsets of conventional T cells: helper T cells which express CD4, and cytotoxic T cells which express CD8. Th1 cells are one of several subsets of helper T cell subsets. Th1 cells promote cell-mediated immunity to combat viral infection, intracellular bacteria, and tumor cells. Both Th1 and cytotoxic T cells can produce IFN-γ and TNF-α.

    Product Usage Information

    Cytokine production by T cells can be induced by activating cells through a variety of methods. These include stimulation through the T cell receptor by incubation with an agonist CD3 antibody such as Human CD3ε Activating (OKT3) Mouse mAb (Low Endotoxin, Azide-free), or treatment with a combination of PMA (phorbol-12-myristate-13-acetate) and Ionomycin. The addition of Brefeldin A and Monensin during the cell stimulation will limit cytokine secretion and facilitate detection of intracellular cytokines. All antibodies in this kit are compatible with the Flow Cytometry TritonTM X-100 Permeabilization Protocol for Directly Conjugated Antibodies and can be used in a single staining mix on fixed and permeabilized cells. Prior to fixation and antibody incubation, we recommend adding a fixable viability dye such as the Ghost Dye™ Violet 510 Viability Dye to enable identification and exclusion of dead cells from analysis.

    Gating strategy for observing production of Th1 cytokines by T cells:
    If a fixable viability dye was used, first gate on viable cells. Next, gate on lymphocytes based on forward scatter and side scatter. To observe cytokine production by helper T cells, gate on the CD3+CD4+ cells within the lymphocyte gate and then identify cells within this gate that are positive for staining of IFN-γ and/or TNF-α. To observe cytokine production by cytotoxic T cells, gate on the CD3+CD8+ cells within the lymphocyte gate and then identify cells within this gate that are positive for staining of IFN-γ and/or TNF-α.

    Storage

    CD3 (UCHT1) Mouse mAb (PE Conjugate), CD4 (RPA-T4) Mouse mAb (violetFluor™ 450 Conjugate), and CD8α (RPA-T8) Mouse mAb (PE-Cy7® Conjugate) are supplied in 10 mM NaH2PO4, 150 mM NaCl, 0.09% NaN3, 0.1% gelatin, pH7.2. TNF-α (D1G2) Rabbit mAb (Alexa Fluor® 647 Conjugate) and IFN-γ (D3H2) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) are supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4oC. Do not aliquot the antibodies. Protect from light. Do not freeze.

    All components in this kit are stable in accordance with the date printed on the outer packaging label when stored at the recommended temperature. Please refer to product labels, datasheets, or web pages for specific "Best By" dates for each individual component.

    Specificity / Sensitivity

    Each antibody in the Human T Cell Th1 Cytokine Response Flow Cytometry Panel detects endogenous levels of its target protein. CD3 (UCHT1) Mouse mAb (PE Conjugate) recognizes CD3ε. CD3 (UCHT1) Mouse mAb (PE Conjugate), CD4 (RPA-T4) Mouse mAb (violetFluor™ 450 Conjugate), and CD8α (RPA-T8) Mouse mAb (PE-Cy7® Conjugate) detect epitopes within the extracellular domains. TNF-α (D1G2) Rabbit mAb (Alexa Fluor® 647 Conjugate) and IFN-γ (D3H2) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) detect epitopes within the intracellular domains.

    Species Reactivity:

    Human

    Source / Purification

    Monoclonal antibodies were purified from tissue culture supernatant via affinity chromatography. The purified antibodies were conjugated under optimal conditions, with unreacted dye removed from the preparation.

    Limited Uses

    Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    For Research Use Only. Not For Use In Diagnostic Procedures.
    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    Cy and CyDye are registered trademarks of GE Healthcare.
    This product is provided under an intellectual property license from Life Technologies Corporation. The transfer of this product is conditioned on the buyer using the purchased product solely in research conducted by the buyer, excluding contract research or any fee for service research, and the buyer must not (1) use this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; or (c) manufacturing or quality assurance or quality control, and/or (2) sell or transfer this product or its components for resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or [email protected].
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