Mouse Activated T Cell Markers Flow Cytometry Panel #62447
Product Information
Protocol
Product Description
The Mouse Activated T Cell Markers Flow Cytometry Panel can be used to identify activated 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. Upon stimulation through the T cell receptor, T cells upregulate expression of CD69 and IL-2Rα/CD25. Regulatory T cells constitutively express IL-2Rα/CD25.
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. Upon stimulation through the T cell receptor, T cells upregulate expression of CD69 and IL-2Rα/CD25. Regulatory T cells constitutively express IL-2Rα/CD25.
Product Usage Information
All antibodies in this kit are compatible with the Flow Cytometry, Live Cell Protocol for Directly Conjugated Antibodies and can be used in a single staining mix. After antibody staining and prior to acquisition on a flow cytometer, we recommend adding a membrane impermeable viability dye such as Propidium Iodide or 7-AAD to enable identification and exclusion of dead cells from the analysis.
Gating strategy for identifying activated T cells:
If Propidium Iodide or 7-AAD was used, first gate on viable cells. Next, gate on lymphocytes based on forward scatter and side scatter. Helper T cells are the CD3+CD4+ cells within the lymphocyte gate. Activated helper T cells are positive for expression of CD69 and/or IL-2Rα/CD25. Regulatory T cells can also express IL-2Rα/CD25. Cytotoxic T cells are the CD3+CD8+ cells within the lymphocyte gate. Activated cytotoxic T cells are positive for expression of CD69 and/or IL-2Rα/CD25.
Gating strategy for identifying activated T cells:
If Propidium Iodide or 7-AAD was used, first gate on viable cells. Next, gate on lymphocytes based on forward scatter and side scatter. Helper T cells are the CD3+CD4+ cells within the lymphocyte gate. Activated helper T cells are positive for expression of CD69 and/or IL-2Rα/CD25. Regulatory T cells can also express IL-2Rα/CD25. Cytotoxic T cells are the CD3+CD8+ cells within the lymphocyte gate. Activated cytotoxic T cells are positive for expression of CD69 and/or IL-2Rα/CD25.
Storage
Antibodies are supplied in 10 mM NaH2PO4, 150 mM NaCl, 0.09% NaN3, 0.1% gelatin, pH7.2. 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 Activated T Cell Markers Flow Cytometry Panel detects endogenous levels of its target protein. CD3 (17A2) Rat mAb (PE Conjugate) recognizes CD3ε, CD3γ, and CD3δ proteins and detects epitopes within the extracellular domains. CD4 (RM4-5) Rat mAb (violetFluor™ 450 Conjugate), CD8α (2.43) Rat mAb (PE-Cy7® Conjugate), IL2-Rα/CD25 (PC61.5) Rat mAb (PerCP-Cy5.5® Conjugate), and CD69 (H1.2F3) Hamster mAb (FITC Conjugate) detect epitopes within the extracellular 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.
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