V450 Rat anti-Mouse CD45R
Clone RA3-6B2 (RUO)
- Brand BD Horizon™
- Alternative Name B220; Ly-5; CD45R; LCA; Ptprc; Protein tyrosine phosphatase receptor type C
- Concentration 0.2 mg/ml
- Isotype Rat IgG2a, κ
- Reactivity Mouse (QC Testing)
Flow cytometry (Routinely Tested)
- Immunogen Mouse Abelson Leukemia Virus-Induced pre-B tumor cells
- Storage Buffer Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The RA3-6B2 monoclonal antibody specifically binds to an epitope on the extracellular domain of the transmembrane CD45 glycoprotein which is dependent upon the expression of exon A and specific carbohydrate residues. It is expressed on B lymphocytes at all stages from pro-B through mature and activated B cell, but it is decreased on plasma cells and a subset of memory B cells. The levels of CD45R expression on the B-cell lineage appear to be developmentally regulated. It is also reportedly found on the abnormal T cells involved in the lymphadenopathy of lpr/lpr and gld/gld mutant mice, on lytically active subsets of lymphokine-activated killer cells (NK cells and non-MHC-restricted CTL), on apoptotic T lymphocytes of mice injected with bacterial superantigen, on a population of NK-cell precursors in the bone marrow, and on B-lymphocyte, T-lymphocyte, and macrophage progenitors in fetal liver. The CD45R antigen has been reported not to be on hematopoietic stem cells, naive T lymphocytes, or MHC-restricted CTL. CD45 is a member of the Protein Tyrosine Phosphatase (PTP) family: Its intracellular (COOH-terminal) region contains two PTP catalytic domains, and the extracellular region is highly variable due to alternative splicing of exons 4, 5, and 6 (designated A, B, and C, respectively), plus differing levels of glycosylation. The CD45 isoforms detected in the mouse are cell type-, maturation, and activation state-specific. The CD45 isoforms play complex roles in T-cell and B-cell antigen receptor signal transduction. CD45R is commonly used as a pan B-cell marker; however, CD19 expression, detectable by the rat anti-mouse CD19 antibody (clone 1D3), is reported to be more restricted to the B-cell lineage. The rat anti-mouse CD45R antibody (clone RA3-6B2) has been reported to enhance isotype switching during in vitro B-cell responses and to inhibit in vivo B-cell responses. Cross-reaction of the RA3-6B2 clone with activated human T lymphocytes has also been reportedly observed.
The antibody is conjugated to BD Horizon™ V450, which has been developed for use in multicolor flow cytometry experiments and is available exclusively from BD Biosciences. It is excited by the Violet laser Ex max of 406 nm and has an Em Max at 450 nm. Conjugates with BD Horizon™ V450 can be used in place of Pacific Blue™ conjugates.
BD Horizon™ V450 is a coumarin dye excited by the violet laser that exhibits spectral properties similar to Pacific Blue™. Conjugates are typically as bright or brighter than comparable reagents conjugated to Pacific Blue™. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon BV421, Pacific Blue™, and BD Horizon V450 cannot be used simultaneously.
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Preparation and Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
The antibody was conjugated with BD Horizon™ V450 under optimum conditions, and unreacted BD Horizon™ V450 was removed.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- BD Horizon V450 has a maximum absorption of 406 nm and maximum emission of 450 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.