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20164
IL2-Rα/CD25 (BC96) Mouse mAb (APC Conjugate)

IL2-Rα/CD25 (BC96) Mouse mAb (APC Conjugate) #20164

APPLICATIONS

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

Flow cytometric analysis of live human peripheral blood mononuclear cells using IL2-Rα/CD25 (BC96) Mouse mAb (APC Conjugate) (right) and co-stained with CD4 (RPA-T4) Mouse mAb (FITC Conjugate) #48705 compared to concentration-matched Mouse (MOPC-21) mAb IgG1 Isotype Control (APC Conjugate) #49083 (left).

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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, 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.

IL2-Rα/CD25 (BC96) Mouse mAb (APC Conjugate) recognizes endogenous levels of total IL2-Rα/CD25 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 allophycocyanin (APC) and tested in-house for direct flow cytometric analysis in human cells.

Interleukin-2 (IL-2) is a T cell stimulatory cytokine best known for inducing T cell proliferation and NK cell proliferation and activation (1,2). IL-2 also promotes peripheral development of regulatory T cells (Tregs) (3,4). Conversely, IL-2 is involved in the activation-induced cell death (AICD) that is observed post T cell expansion by increasing levels of Fas on CD4+ T cells (5). The effects of IL-2 are mediated through a trimeric receptor complex consisting of IL-2Rα, IL-2Rβ, and the common gamma chain, γc (1,2). IL-2Rα binds exclusively to IL-2 with low affinity and increases the binding affinity of the whole receptor complex including IL-2Rβ and γc subunits. IL-15 also binds to IL-2Rβ (1,2). γc is used by other cytokines including IL-4, IL-7, IL-9, IL-15, and IL-21 (1,2). Binding of IL-2 initiates signaling cascades involving Jak1, Jak3, Stat5, and the PI3K/Akt pathways (1,2).

  1. Ma, A. et al. (2006) Annu Rev Immunol 24, 657-79.
  2. Gaffen, S.L. and Liu, K.D. (2004) Cytokine 28, 109-23.
  3. Fehérvari, Z. et al. (2006) Trends Immunol 27, 109-11.
  4. Antony, P.A. et al. (2006) J Immunol 176, 5255-66.
  5. Jaleco, S. et al. (2003) J Immunol 171, 61-8.
Entrez-Gene Id
3559
Swiss-Prot Acc.
P01589
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

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To Purchase # 20164S

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Product # Size Price
20164S
500 µl (100 tests) $ 159.0