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35094
CD127/IL-7Rα (A7R34) Rat mAb (PerCP-Cy5.5® Conjugate)
Antibody Conjugates

CD127/IL-7Rα (A7R34) Rat mAb (PerCP-Cy5.5® Conjugate) #35094

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Flow cytometric analysis of live mouse splenocytes using CD127/IL-7Rα (A7R34) Rat mAb (PerCP-Cy5.5® Conjugate) and co-stained with CD3 (17A2) Rat mAb (APC Conjugate) #24265 (right), compared to concentration-matched Rat Isotype Control (PerCP-Cy5.5® Conjugate) (left).

To Purchase # 35094S
Product # Size Price
35094S
100 µg $ 229

Supporting Data

REACTIVITY M
SENSITIVITY Endogenous
MW (kDa)
Isotype Rat IgG2a, kappa

Product Description

This Cell Signaling Technology antibody is conjugated to PerCP-Cy5.5® and tested in-house for direct flow cytometric analysis in mouse cells.

Product Usage Information

For optimal flow cytometry results, we recommend 0.25 μg of antibody per test.

Application Dilutions
Flow Cytometry 1:80

Storage:

Supplied in 10 mM NaH2PO4, 150 mM NaCl, 0.09% NaN3, 0.1% gelatin, pH 7.2. This product is stable for 6 months when stored at 4ºC. Do not aliquot the antibody. Protect from light. Do not freeze.

Protocol

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Flow Cytometry

A. Solutions and Reagents

NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.

  1. 20X Phosphate Buffered Saline (PBS): (#9808) To prepare 1 L 1X PBS: add 50 ml 20X PBS to 950 ml dH2O, mix.
  2. Incubation Buffer: Dissolve 0.5 g Bovine Serum Albumin (BSA) (#9998) in 100 ml 1X PBS. Store at 4°C.

B. Immunostaining

  1. Count cells using a hemacytometer or alternative method.
  2. Aliquot 0.5-1x106 cells into each assay tube (by volume).
  3. Add 2-3 ml Incubation Buffer to each tube and rinse by centrifugation. Repeat.
  4. Resuspend cells in 100 µl of diluted primary antibody (prepared in incubation buffer at the recommended dilution).
  5. Incubate for 1 hr at room temperature.
  6. Wash by centrifugation in 2-3 ml incubation buffer.
  7. Resuspend cells in the appropriate volume of PBS and analyze on flow cytometer.

posted June 2017

Protocol Id: 1504

Specificity / Sensitivity

CD127/IL-7Rα (A7R34) Rat mAb (PerCP-Cy5.5® Conjugate) recognizes endogenous levels of total CD127/IL-7Rα protein. This antibody detects an epitope within the extracellular domain.

Species Reactivity:

Mouse

Source / Purification

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.

Background

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).

  1. Rochman, Y. et al. (2009) Nat Rev Immunol 9, 480-90.
  2. Levin, S.D. et al. (1999) J Immunol 162, 677-83.
  3. Schluns, K.S. et al. (2000) Nat Immunol 1, 426-32.
  4. Rochman, Y. and Leonard, W.J. (2008) J Immunol 181, 7699-705.
  5. McElroy, C.A. et al. (2012) Proc Natl Acad Sci U S A 109, 2503-8.
  6. Sudo, T. et al. (1993) Proc Natl Acad Sci U S A 90, 9125-9.

Pathways & Proteins

Explore pathways + proteins related to this product.

For Research Use Only. Not For Use In Diagnostic Procedures.

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