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47972
Mouse Essential T Cell Markers Flow Cytometry Panel
Flow Cytometry Kits & Reagents
Assay Kit

Mouse Essential T Cell Markers Flow Cytometry Panel #47972

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Flow cytometric analysis of live mouse splenocytes using CD4 (RM4-5) Rat mAb (violetFluor 450 Conjugate)
(solid line) compared to concentration-matched Rat Isotype Control (violetFluor 450 Conjugate) (dashed line).
Flow cytometric analysis of live mouse splenocytes using FoxP3 (3G3) Mouse mAb (APC Conjugate) (right) compared to a concentration-matched Mouse (MOPC-21) mAb IgG1 Isotype Control (APC Conjugate) #49083 (left), co-stained with CD4 (RM4-5) Rat mAb (FITC Conjugate) #96127.
Flow cytometric analysis of live mouse splenocytes using CD8α (2.43) Rat mAb (PE-Cy7® Conjugate) (solid line) compared to concentration-matched Rat (LTF-2) mAb IgG2b Isotype Control (PE-Cy7® Conjugate) #43426 (dashed line).
Flow cytometric analysis of human peripheral blood mononuclear cells, untreated (left column) or treated with anti-CD3 plus anti-CD28 (10 μg/ml, 72 hr; right column), using Granzyme B (D2H2F) Rabbit mAb (Alexa Fluor® 488 Conjugate) (top row) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 488 Conjugate) #2975 (bottom row), and co-stained with a CD8 Antibody.
Flow cytometric analysis of live mouse splenocytes using CD3 (17A2) Rat mAb (PE Conjugate) (solid line) compared to concentration-matched Rat (LTF-2) mAb IgG2b Isotype Control (PE Conjugate) #27426 (dashed line).
To Purchase # 47972
Cat. # Size Qty. Price
47972S
1 Kit  (50 assays)

Important Ordering Details

Custom Ordering Details: Product is assembled upon order. Please allow up to three business days for your product to be processed.

Protocol

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Flow Cytometry using the FoxP3/Transcription Factor Fixation/Permeabilization Kit

This antibody has been validated for Flow Cytometry using the FoxP3/Transcription Factor Fixation/Permeabilization Kit #43481. Please refer to the protocol provided with the kit for detailed instructions.

Protocol Id: 1804

Product Description

The Mouse Essential T Cell Markers Flow Cytometry Panel can be used to identify major subsets of mouse T cells.

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. Regulatory CD4+ T cells express FoxP3. Upon antigen stimulation, T cells upregulate expression of Granzyme B, a serine protease that is secreted by these cells to mediate target cell cytotoxicity.

Product Usage Information

All antibodies in this kit are compatible with the FoxP3/Transcription Factor Fixation/Permeabilization Kit #43481 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 identifying essential T cell subsets:
If a fixable viability dye was used, first gate on viable cells. Next, gate on lymphocytes based on forward scatter and side scatter. Conventional helper T cells are the CD3+CD4+ cells within the lymphocyte gate. Regulatory CD3+CD4+ T cells are positive for expression of FoxP3. Conventional cytotoxic T cells are the CD3+CD8+ cells within the lymphocyte gate. Both CD3+CD4+ and CD3+CD8+ cells can be positive for expression of the cytotoxic molecule Granzyme B.

Storage

CD3 (17A2) Rat mAb (PE Conjugate), CD4 (RM4-5) Rat mAb (violetFluor 450 Conjugate), FoxP3 (3G3) Mouse mAb (APC Conjugate), and CD8α (2.43) Rat mAb (PE-Cy7® Conjugate) are supplied in 10 mM NaH2PO4, 150 mM NaCl, 0.09% NaN3, 0.1% gelatin, pH7.2. Granzyme B (D2H2F) Rabbit mAb (Alexa Fluor® 488 Conjugate) is 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 Mouse Essential T Cell Markers Flow Cytometry Panel detects endogenous levels of its target protein. CD3 (17A2) Rat mAb (PE Conjugate), CD4 (RM4-5) Rat mAb (violetFluor 450 Conjugate), and CD8α (2.43) Rat mAb (PE-Cy7® Conjugate) detect epitopes within the extracellular domains. FoxP3 (3G3) Mouse mAb (APC Conjugate) and Granzyme B (D2H2F) Rabbit mAb (Alexa Fluor® 488 Conjugate) detect epitopes within the intracellular domains.

Species Reactivity:

Mouse

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

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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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