PerCP Rat Anti-Mouse CD45R/B220
Clone RA3-6B2 (RUO)
- Brand BD Pharmingen™
- Concentration 0.2 mg/ml
- Isotype Rat IgG2a, κ
- Reactivity Mouse (QC Testing) Human (Reported)
Flow cytometry (Routinely Tested)
- Immunogen Mouse Abelson Leukemia Virus-Induced pre-B tumor cells
- Storage Buffer Aqueous buffered solution containing ≤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.
PerCP is a component of the photosynthetic apparatus found in the dinoflagellate Glenodinium. PerCP is a protein complex with a molecular weight of approximately 35 kDa. Due to its photobleaching characteristics, PerCP conjugates are not recommended for use on flow cytometers with high-power lasers (>25 mW).
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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 PerCP under optimum conditions, and unconjugated antibody and free PerCP were removed. Storage of PerCP conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.
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/us/s/resources for technical protocols.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- PerCP is a photosynthetic accessory pigment from Glenodinium species of dinoflagellates, which is excited by the 488-nm light of an Argon ion laser and fluoresces at 675 nm. Therefore, PerCP-labelled antibodies can be used with FITC- and R-PE–labelled reagents in most single-laser flow cytometers with no significant spectral overlap of PerCP fluorescence with that of FITC or R-PE. PerCP has been reported to undergo significant photobleaching, the magnitude of which increases as laser power is increased or beam focus is narrowed. For third-color flow¬cytometric analysis using ≥25-mW laser power, we recommend PE-Cy5-, PE-Cy7–, or PerCP-Cy5.5-conjugated reagents.
- 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.