North America
South America
Australia & New Zealand
Europe
Middle East / Africa
Asia / Pacific
×
×
Pacific Blue™ Rat Anti-Mouse CD45R RA3-6B2 RUO

Pacific Blue™ Rat Anti-Mouse CD45R 

Clone  RA3-6B2   (RUO)

  • Brand BD Pharmingen™
  • 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)  Human (Reported) 
  • Application 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
Product Details Recommended Assay References

Description

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.

Format
  • Format Pacific Blue™
  • Excitation Source Violet 405 nm
  • Excitation Max 401 nm
  • Emission Max 452 nm

Pacific Blue™ is based on the 6,8-difluoro-7-hydroxycoumarin fluorophore, and is strongly fluorescent, even at neutral pH. 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.

Other Formats →

Suggested Companion Products

Pacific Blue™ Rat IgG2a, κ Isotype Control RUO
0.1 mg Cat No: 558109

Stain Buffer (FBS) RUO
500 mL Cat No: 554656

Stain Buffer (BSA) RUO
500 mL Cat No: 554657

Resources & Tools
 Spectrum Viewer  Panel Designer  Spectrum Viewer  Download TDS  Regulatory Document Website
Preparation and Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody is conjugated to Pacific Blue™ under optimum conditions, and unreacted Pacific Blue™ was removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  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. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  5. Pacific Blue™ has a maximum absorption of 416 nm and maximum emission of 451 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.


  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: Flow cytometry). 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: Flow cytometry). View reference

  3. Ballas ZK, Rasmussen W. Lymphokine-activated killer cells. VII. IL-4 induces an NK1.1+CD8 alpha+beta- TCR-alpha beta B220+ lymphokine-activated killer subset. J Immunol. 1993; 150(1):17-30. (Biology: Flow cytometry). View reference

  4. Bleesing JJ, Morrow MR, Uzel G, Fleisher TA. Human T cell activation induces the expression of a novel CD45 isoform that is analogous to murine B220 and is associated with altered O-glycan synthesis and onset of apoptosis. Cell Immunol. 2001; 213(1):72-81. (Biology: Flow cytometry). View reference

  5. Coffman RL. Surface antigen expression and immunoglobulin gene rearrangement during mouse pre-B cell development. Immunol Rev. 1982; 69:5-23. (Immunogen: Flow cytometry). View reference

  6. 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: Flow cytometry). View reference

  7. Driver DJ, McHeyzer-Williams LJ, Cool M, Stetson DB, McHeyzer-Williams MG. Development and maintenance of a B220- memory B cell compartment. J Immunol. 2001; 167(3):1393-1405. (Biology: Flow cytometry). View reference

  8. 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: Flow cytometry). View reference

  9. 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: Flow cytometry). View reference

  10. 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: Flow cytometry). View reference

  11. 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.

  12. Kobata T, Takasaki K, Asahara H, et al. Apoptosis with FasL+ cell infiltration in the periphery and thymus of corrected autoimmune mice. Immunology. 1997; 92(2):206-213. (Biology: Flow cytometry). View reference

  13. Krop I, de Fougerolles AR, Hardy RR, Allison M, Schlissel MS, Fearon DT. Self-renewal of B-1 lymphocytes is dependent on CD19. Eur J Immunol. 1996; 26(1):238-242. (Biology: Flow cytometry). View reference

  14. Laouar Y, Ezine S. In vivo CD4+ lymph node T cells from lpr mice generate CD4-CD8-B220+TCR-beta low cells. J Immunol. 1994; 153(9):3948-3955. (Biology: Flow cytometry). View reference

  15. Puzanov IJ, Bennett M, Kumar V. IL-15 can substitute for the marrow microenvironment in the differentiation of natural killer cells. J Immunol. 1996; 157(10):4282-4285. (Biology: Flow cytometry). View reference

  16. Renno T, Hahne M, Tschopp J, MacDonald HR. Peripheral T cells undergoing superantigen-induced apoptosis in vivo express B220 and upregulate Fas and Fas ligand. J Exp Med. 1996; 183(2):431-437. (Biology: Flow cytometry). View reference

  17. Rolink A, ten Boekel E, Melchers F, Fearon DT, Krop I, Andersson J. A subpopulation of B220+ cells in murine bone marrow does not express CD19 and contains natural killer cell progenitors. J Exp Med. 1996; 183(1):187-194. (Biology: Flow cytometry). View reference

  18. Sagara S, Sugaya K, Tokoro Y, et al. B220 expression by T lymphoid progenitor cells in mouse fetal liver. J Immunol. 1997; 158(2):666-676. (Biology: Flow cytometry). View reference

558108 Rev. 3
Instruments
Reagents
Applications
Sales & Support
BD BioSciences