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Purified Rat Anti-Mouse CD45R
Purified Rat Anti-Mouse CD45R

Immunohistochemical staining of CD45R.

A formalin-fixed paraffin-embedded section from normal mouse spleen was stained with the purified Rat Anti-Mouse CD45R antibody.  CD45R+ B-lymphocytes can be identified by the brown labeling of the cell membranes (20X magnification).

Immunohistochemical staining of CD45R.

A formalin-fixed paraffin-embedded section from normal mouse spleen was stained with the purified Rat Anti-Mouse CD45R antibody.  CD45R+ B-lymphocytes can be identified by the brown labeling of the cell membranes (20X magnification).

Product Details
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BD Pharmingen™
B220; Ly-5; CD45R; LCA; Ptprc; Protein tyrosine phosphatase receptor type C
Mouse (QC Testing)
Rat IgG2a, κ
Mouse Abelson Leukemia Virus-Induced pre-B tumor cells
Flow cytometry (Routinely Tested), Immunohistochemistry-frozen, Immunohistochemistry-paraffin (Tested During Development)
31.25 µg/ml
AB_393581
Aqueous buffered solution containing BSA, goat serum, and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Recommended Assay Procedures

Immunohistochemistry: This antibody may be used for the immunohistochemical staining of formalin-fixed paraffin-embedded sections of mouse spleen or thymus.  No pretreatment for antigen retrieval is required. This antibody has been reported to stain for all B cells but not plasma cells. The isotype control recommended for use with this antibody is purified rat IgG2a (Cat. No. 559073). For optimal indirect immunohistochemical staining, this antibody should be titrated (1:10 to 1:50 dilution) and visualized via a three-step staining procedure in combination with biotinylated polyclonal anti-rat Ig (Cat. No. 559286) as the secondary antibody and Streptavidin-HRP (Cat. No. 550946) together with the DAB detection system (Cat. No. 550880).  More conveniently, the anti-rat Ig HRP detection kit (Cat. No. 551013) can be used which contains the biotinylated secondary antibody, the antibody diluent, Streptavidin-HRP and DAB substrates for use in the staining procedure.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. 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.
  4. This antibody has been developed for the immunohistochemistry application. However, a routine immunohistochemistry test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
  5. An isotype control should be used at the same concentration as the antibody of interest.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
550286 Rev. 3
Antibody Details
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RA3-6B2

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.

550286 Rev. 3
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
550286 Rev.3
Citations & References
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Development References (8)

  1. Allman DM, Ferguson SE, Cancro MP. Peripheral B cell maturation. I. Immature peripheral B cells in adults are heat-stable antigenhi and exhibit unique signaling characteristics. J Immunol. 1992; 149(8):2533-2540. (Biology). View Reference
  2. Asensi V, Kimeno K, Kawamura I, Sakumoto M, Nomoto K. Treatment of autoimmune MRL/lpr mice with anti-B220 monoclonal antibody reduces the level of anti-DNA antibodies and lymphadenopathies. Immunology. 1989; 68(2):204-208. (Biology). View Reference
  3. Coffman RL. Surface antigen expression and immunoglobulin gene rearrangement during mouse pre-B cell development. Immunol Rev. 1982; 69:5-23. (Immunogen: Immunoprecipitation). View Reference
  4. Domiati-Saad R, Ogle EW, Justement LB. Administration of anti-CD45 mAb specific for a B cell-restricted epitope abrogates the B cell response to a T-dependent antigen in vivo. J Immunol. 1993; 151(11):5936-5947. (Biology). View Reference
  5. George A, Rath S, Shroff KE, Wang M, Durdik JM. Ligation of CD45 on B cells can facilitate production of secondary Ig isotypes. J Immunol. 1994; 152(3):1014-1021. (Biology). View Reference
  6. Hardy RR, Carmack CE, Shinton SA, Kemp JD, Hayakawa K. Resolution and characterization of pro-B and pre-pro-B cell stages in normal mouse bone marrow. J Exp Med. 1991; 173(5):1213-1225. (Biology). View Reference
  7. Hathcock KS, Hirano H, Murakami S, Hodes RJ. CD45 expression by B cells. Expression of different CD45 isoforms by subpopulations of activated B cells. J Immunol. 1992; 149(7):2286-2294. (Biology). View Reference
  8. Johnson P, Maiti A, Ng DHW. CD45: A family of leukocyte-specific cell surface glycoproteins. In: Herzenberg LA, Weir DM, Herzenberg LA, Blackwell C , ed. Weir's Handbook of Experimental Immunology, Vol 2. Cambridge: Blackwell Science; 1997:62.1-62.16.
View All (8) View Less
550286 Rev. 3

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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.