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93204
CD127/IL-7Rα (A7R34) Rat mAb (PE-Cy7® Conjugate)
Antibody Conjugates

CD127/IL-7Rα (A7R34) Rat mAb (PE-Cy7® Conjugate) #93204

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Flow cytometric analysis of live mouse splenocytes using CD127/IL-7Rα (A7R34) Rat mAb (PE-Cy7® Conjugate) and co-stained with CD3 (17A2) Rat mAb (APC Conjugate) #24265 (right), compared to concentration-matched Rat Isotype Control (PE-Cy7® Conjugate) (left).

To Purchase # 93204S
Product # Size Price
93204S
100 µg $ 299

Supporting Data

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

Application Key:

  • W-Western
  • IP-Immunoprecipitation
  • IHC-Immunohistochemistry
  • ChIP-Chromatin Immunoprecipitation
  • IF-Immunofluorescence
  • F-Flow Cytometry
  • E-P-ELISA-Peptide

Species Cross-Reactivity Key:

  • H-Human
  • M-Mouse
  • R-Rat
  • Hm-Hamster
  • Mk-Monkey
  • Mi-Mink
  • C-Chicken
  • Dm-D. melanogaster
  • X-Xenopus
  • Z-Zebrafish
  • B-Bovine
  • Dg-Dog
  • Pg-Pig
  • Sc-S. cerevisiae
  • Ce-C. elegans
  • Hr-Horse
  • All-All Species Expected

Product Description

This Cell Signaling Technology antibody is conjugated to PE-Cy7® and tested in-house for direct flow cytometric analysis in mouse cells.

Product Usage Information

For optimal flow cytometry results, we recommend 0.5 μg of antibody per test.

Application Dilutions
Flow Cytometry 1:40

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, Live Cell Protocol for Directly Conjugated Antibodies

A. Solutions and Reagents

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

  1. 1X Phosphate Buffered Saline (PBS): To prepare 1 L 1X PBS: add 100 ml 10X PBS (#12528) to 900 ml water, mix.
  2. Antibody Dilution Buffer: Purchase ready-to-use Flow Cytometry Antibody Dilution Buffer (#13616), or prepare a 0.5% BSA PBS buffer by dissolving 0.5 g Bovine Serum Albumin (BSA) (#9998) in 100 ml 1X PBS. Store at 4°C.

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/flowdyes for a listing of cellular dyes validated for use in flow cytometry.

B. Immunostaining

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.

  1. Aliquot desired number of cells into tubes or wells. (Generally, 5x105 to 1x106 cells per assay.)
  2. Pellet cells by centrifugation and remove supernatant.
  3. Resuspend cells in 100 µl of diluted primary antibody, prepared in Antibody Dilution Buffer at a recommended dilution or as determined via titration.
  4. Incubate for 30 min to 1 hr on ice. Protect from light.
  5. Wash by centrifugation in Antibody Dilution Buffer. Discard supernatant. Repeat.
  6. Resuspend cells in 200-500 µl of Antibody Dilution Buffer and analyze on flow cytometer.

posted June 2017

revised August 2019

Protocol Id: 1504

Specificity / Sensitivity

CD127/IL-7Rα (A7R34) Rat mAb (PE-Cy7® 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.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Cy and CyDye are registered trademarks of GE Healthcare.