|H M||Endogenous||Rat IgG2a, kappa|
Flow cytometric analysis of live mouse splenocytes using CD45R/B220 (RA3-6B2) Rat mAb (violetFluor™ 450 Conjugate)
(solid line) compared to concentration-matched Rat Isotype Control (violetFluor™ 450 Conjugate) (dashed line).Learn more about how we get our images.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
posted June 2017
Protocol Id: 1504
For optimal flow cytometry results, we recommend 0.5 μg of antibody per test.
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.
CD45R/B220 (RA3-6B2) Rat mAb (violetFluor™ 450 Conjugate) recognizes endogenous levels of total CD45R/B220 protein. This antibody detects an epitope within the extracellular domain.
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 violetFluor™ 450 and tested in-house for direct flow cytometric analysis in mouse cells.
The protein phosphatase (PTP) receptor CD45 is a type I transmembrane protein comprised of a pair of intracellular tyrosine phosphatase domains and a variable extracellular domain generated by alternative splicing (1). The catalytic activity of CD45 is a function of the first phosphatase domain (D1) while the second phosphatase domain (D2) may interact with and stabilize the first domain, or recruit/bind substrates (2,3). CD45 interacts directly with antigen receptor complex proteins or activates Src family kinases involved in the regulation of T- and B-cell antigen receptor signaling (1). Specifically, CD45 dephosphorylates Src-family kinases Lck and Fyn at their conserved negative regulatory carboxy-terminal tyrosine residues and upregulates kinase activity. Conversely, studies indicate that CD45 can also inhibit Lck and Fyn by dephosphorylating their positive regulatory autophosphorylation site. CD45 appears to be both a positive and a negative regulator that conducts signals depending on specific stimuli and cell type (1). Human leukocytes including lymphocytes, eosinophils, monocytes, basophils, and neutrophils express CD45, while erythrocytes and platelets are negative for CD45 expression (4).
The RA3-6B2 clone is raised against an isoform of CD45 known as CD45R/B220. CD45R/B220 is widely used as a marker for B cells, T cell subsets, and NK cell subsets in both human and mouse (5).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. violetFluor is a registered trademark of Tonbo Biosciences.
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