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86135
CD8α (RPA-T8) Mouse mAb (redFluor™ 710 Conjugate)

CD8α (RPA-T8) Mouse mAb (redFluor™ 710 Conjugate) #86135

APPLICATIONS

REACTIVITY SENSITIVITY MW (kDa) Isotype
H Endogenous Mouse IgG1
Flow Cytometry

Flow cytometric analysis of live human peripheral blood mononuclear cells using CD8α (RPA-T8) Mouse mAb (redFluor™ 710 Conjugate) (solid line) compared to concentration-matched Mouse (MOPC-21) mAb IgG1 Isotype Control (redFluor™ 710 Conjugate) #35935 (dashed line).

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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

Application Dilutions
Flow Cytometry 1:20
Storage:

Supplied in 10 mM NaH2PO4, 150 mM NaCl, 0.09% NaN3, 0.1% gelatin, pH7.2. This product is stable for 6 months when stored at 4ºC. Do not aliquot the antibody. Protect from light. Do not freeze.

CD8α (RPA-T8) Mouse mAb (redFluor™ 710 Conjugate) recognizes endogenous levels of total CD8α protein. This antibody detects an epitope within the extracellular domain.

Species Reactivity:

Human

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.

This Cell Signaling Technology antibody is conjugated to redFluor™ 710 and tested in-house for direct flow cytometric analysis in human cells.

Cluster of Differentiation 8 (CD8) is a disulphide-linked heterodimer consisting of the unrelated α and β subunits. Each subunit is a glycoprotein composed of a single extracellular Ig-like domain, a polypeptide linker, a transmembrane part and a short cytoplasmic tail. On T cells, CD8 is the coreceptor for the T cell receptor (TCR), and these two distinct structures recognize the Antigen–Major Histocompatibility Complex (MHC). Specifically, the Ig-like domain of CD8α interacts with the α3-domain of the MHC class I molecule. CD8 ensures specificity of the TCR–antigen interaction, prolongs the contact between the T cell and the antigen presenting cell, and the α chain recruits the tyrosine kinase Lck, which is essential for T cell activation (1).

The RPA-T8 antibody is widely used as a phenotypic marker for CD8 on cytotoxic T cells and thymocytes (2,3), as well as on certain cell types that do not express the TCR, including some NK cells (4).

  1. Zamoyska, R. (1994) Immunity 1, 243-246.
  2. Friberg, H. et al. (2011) Immunol Cell Biol 89, 122-9.
  3. Reissfelder, C. et al. (2015) J Clin Invest 125, 739-51.
  4. Addison, E.G. et al. (2005) Immunology 116, 354-61.
Entrez-Gene Id
925
Swiss-Prot Acc.
P01732
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
redFluor is a registered trademark of Tonbo Biosciences.

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To Purchase # 86135S

Product Number Size Price
86135S 500 µl (100 tests) $319.00.0
Quantity Subtotal
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