Mouse Essential T Cell Markers Flow Cytometry Panel #47972
Important Ordering Details
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Protocol
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.
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.
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.
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.
限制使用
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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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