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Safety Data Sheet (SDS) Brilliant Violet 421 Anti-mouse/human CD45R/B220 Antibody     Product Data Sheet (PDF)    
Brilliant Violet 421™ Anti-mouse/human CD45R/B220 Antibody
1116195 125 µl $150.00       
1116255 50 µg $235.00       
1116200 500 µl $345.00       
Clone: RA3-6B2
Isotype: Rat IgG2a, κ
Reactivity: Mouse, Human, Cross-Reactivity: Cat (Feline)
Immunogen: Abelson murine leukemia virus-induced pre-B tumor cells
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA).
Preparation: The antibody was purified by affinity chromatography and conjugated with Brilliant Violet 421™ under optimal conditions. The solution is free of unconjugated Brilliant Violet 421™ and unconjugated antibody.
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application:

FC - Quality tested
IHC - Validated

Application Notes:

Clone RA3-6B2 has been described to react with an epitope on the extracellular domain of the transmembrane CD45 glycoprotein which is dependent upon the expression of exon A and specific carbohydrate residues. Additional reported applications (for the relevant formats) include: immunoprecipitation1, in vitro and in vivo modulation of B cell responses2-4, and immunohistochemistry of acetone-fixed frozen sections and formalin-fixed paraffin-embedded sections5,6.

Recommended Usage:

Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is ≤5 µl per million cells or 5 µl per 100 µl of whole blood. For immunohistochemistry, a concentration range of 2.5 - 10 µg per ml. It is recommended that the reagent be titrated for optimal performance for each application.

Brilliant Violet 421™ excites at 405 nm and emits at 421 nm. The standard bandpass filter 450/50 nm is recommended for detection. Brilliant Violet 421™ is a trademark of Sirigen Group Ltd.

This product is subject to proprietary rights of Sirigen Inc. and is made and sold under license from Sirigen Inc. The purchase of this product conveys to the buyer a non-transferable right to use the purchased product for research purposes only. This product may not be resold or incorporated in any manner into another product for resale. Any use for therapeutics or diagnostics is strictly prohibited. This product is covered by U.S. Patent(s), pending patent applications and foreign equivalents.

Application References:

1. Coffman RL. 1982. Immunol. Rev. 69:5. (IP)
2. George A, et al. 1994. J. Immunol. 152:1014. (Activ)
3. Asensi V, et al. 1989. Immunology 68:204. (Activ)
4. Domiati-Saad R, et al. 1993. J. Immunol. 151:5936. (Activ)
5. Hata H, et al. 2004. J. Clin. Invest. 114:582. (IHC)
6. Monteith CE, et al. 1996. Can. J. Vet. Res. 60:193. (IHC)
7. Shih FF, et al. 2006. J. Immunol. 176:3438. (FC)
8. Chang C L-T, et al. 2007. J. Immunol. 178:6984.
9. Fazilleau N, et al. 2007. Nature Immunol. 8:753.
10. Lang GL, et al. 2008. Blood 111:2158. PubMed
11. Charles N, et al. 2010. Nat. Med. 16:701. (FC) PubMed
12. del Rio ML, et al. 2011. Transpl. Int. 24:501. (FC) PubMed
13. Murakami R, et al. 2013. PLoS One. 8:73270. PubMed

C57BL/6 mouse splenocytes were stained

C57BL/6 mouse splenocytes were stained with CD45R/B220 (clone RA3-6B2) Brilliant Violet 421™ (filled histogram) or rat IgG2a, κ Brilliant Violet 421™ isotype control (open histogram).

C57BL/6 mouse frozen spleen sections

C57BL/6 mouse frozen spleen sections were fixed with 4% paraformaldehyde (PFA) for ten minutes at room temperature and blocked with 5% FBS plus 5% rat/mouse serum for 30 minutes at room temperature. Then the section was stained with 1.25 µg/ml of anti-mouse/human CD45R/B220 (clone RA3-6B2) Brilliant Violet 421™ (blue), anti-mouse CD5 (clone 53-7.3) Alexa Fluor® 594 (red), and anti-mouse F4/80 (clone BM8) Alexa Fluor® 647 (green) overnight at 4°C. The image was captured with a 10X objective.



Description:

CD45R, also known as B220, is an isoform of CD45. It is a member of the protein tyrosine phosphatase (PTP) family with a molecular weight of approximately 180-240 kD. CD45R is expressed on B cells (at all developmental stages from pro-B cells through mature B cells), activated B cells, and subsets of T and NK cells. CD45R (B220) is also expressed on a subset of abnormal T cells involved in the pathogenesis of systemic autoimmunity in MRL-Faslpr and MRL-Fasgld mice. It plays a critical role in TCR and BCR signaling. The primary ligands for CD45 are galectin-1, CD2, CD3, and CD4. CD45R is commonly used as a pan-B cell marker; however, CD19 may be more appropriate for B cell specificity.

Other Names: B220
Structure: Protein tyrosine phosphatase (PTP) family, 180-240 kD
Distribution: B cells, T cell subset, NK cell subset
Function: Phosphatase, T and B cell activation
Ligand Receptor: Galectin-1, CD2, CD3, CD4
Antigen References:

1. Barclay A, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Trowbridge IS, et al. 1993. Annu. Rev. Immunol. 12:85.
3. Kishihara K, et al. 1993. Cell 74:143.
4. Pulido R, et al. 1988. J. Immunol. 140:3851.