Flow cytometric analysis of live mouse splenocytes using CD127/IL-7Rα (A7R34) Rat mAb (APC Conjugate) and co-stained with CD3 (17A2) Rat mAb (FITC Conjugate) #86603 (right), compared to concentration-matched Rat Isotype Control (APC Conjugate) (left).
|Source/Isotype||Rat IgG2a kappa|
This Cell Signaling Technology antibody is conjugated to APC and tested in-house for direct flow cytometric analysis in mouse cells.
For optimal flow cytometry results, we recommend 0.25 μg of antibody per test.
Supplied in 10 mM NaH2PO4, 150 mM NaCl, 0.09% NaN3, 0.1% gelatin, pH 7.2. Do not aliquot the antibody. Protect from light. Do not freeze.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: When including fluorescent cellular dyes in your experiment (including viability dyes, DNA dyes, etc.), please refer to the dye product page for the recommended protocol. Visit www.cellsignal.com for a full listing of cellular dyes validated for use in flow cytometry.
NOTE: Count cells using a hemocytometer or alternative method.
NOTE: If using whole blood, lyse red blood cells and wash by centrifugation prior to Immunostaining.
NOTE: Optimal centrifugation conditions will vary depending upon cell type and reagent volume. Generally, 150-300g for 1-5 minutes will be sufficient to pellet the cells.
posted June 2017
revised June 2020
Protocol Id: 1504
CD127/IL-7Rα (A7R34) Rat mAb (APC Conjugate) recognizes endogenous levels of total CD127/IL-7Rα protein. This antibody detects an epitope within the extracellular domain.
This monoclonal antibody was purified from tissue culture supernatant via affinity chromatography. The purified antibody was conjugated under optimal conditions, with unreacted dye removed from the preparation.
The IL-7 receptor (IL-7R) is comprised of two protein subunits, CD127/IL-7Rα (IL-7Ralpha) and the common gamma chain (CD132), which is the major signaling component for several cytokines including IL-2, IL-4, IL-9, IL-15, and IL-21 (1). CD127/IL-7Rα is a transmembrane protein belonging to the cytokine receptor homology class 1 (CRH1) and is expressed by a wide variety of cells including immature B cells, thymic natural killer cells, bone marrow stromal cells, and T cells (5-6). On its own, CD127/IL-7Rα functions as a receptor for two cytokine receptor complex signaling cascades: IL-7 and thymic stromal lymphopoietin (TSLP) (2). IL-7 signaling contributes to T cell development and homeostasis whereas TSLP receptor signaling contributes to dendritic cell activation and B cell development. IL-7 signaling is an essential component in regulating the homeostasis of naive and memory T cells as differential expression of CD127/IL-7Rα is observed on naive and activated T cells, which occurs following TCR activation. Specifically, CD127/IL-7Rα expression is downregulated on activated T cells and the subsequent re-expression of CD127/IL-7Rα on these cells is indicative of cells that will differentiate into memory T cells (3-4).
The A7R34 antibody is widely used to identify CD127/IL-7Rα expression on both B and T cells (6).
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