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82116
CD4 (RM4-5) Rat mAb (APC Conjugate)
Antibody Conjugates

CD4 (RM4-5) Rat mAb (APC Conjugate) #82116

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APPLICATIONS

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

Flow cytometric analysis of live mouse splenocytes using CD4 (RM4-5) Rat mAb (APC Conjugate) (solid line) compared to concentration-matched Rat Isotype Control (APC Conjugate) (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

For optimal flow cytometry results, we recommend 0.125 μg of antibody per test.

Application Dilutions
Flow Cytometry 1:160
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.

CD4 (RM4-5) Rat mAb (APC Conjugate) recognizes endogenous levels of total CD4 protein. This antibody detects an epitope within the extracellular domain.

Species Reactivity:

Mouse

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 mouse cells.

Cluster of Differentiation 4 (CD4) is a glycoprotein composed of an amino-terminal extracellular domain (four domains: D1-D4 with Ig-like structures), a transmembrane part and a short cytoplasmic tail. CD4 is expressed on the surface of T helper cells, regulatory T cells, monocytes, macrophages and dendritic cells, and plays an important role in the development and activation of T cells. On T cells, CD4 is the co-receptor for the T cell receptor (TCR), and these two distinct structures recognize the Antigen–Major Histocompatibility Complex (MHC). Specifically, the D1 domain of CD4 interacts with the β2-domain of the MHC class II molecule. CD4 ensures specificity of the TCR–antigen interaction, prolongs the contact between the T cell and the antigen presenting cell and recruits the tyrosine kinase Lck, which is essential for T cell activation (1).

  1. Zamoyska, R. (1994) Immunity 1, 243-6.
Entrez-Gene Id
12504
Swiss-Prot Acc.
P06332
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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Product # Size Price
82116S		 100 µg $ 165

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