Phosphate-buffered solution, pH 7.2, containing no preservative. 0.2 μm filter sterilized. Endotoxin level is < 0.1 EU/μg of the protein (< 0.01 ng/μg of the protein) as determined by the LAL test.
Preparation:
The LEAF™ (Low Endotoxin, Azide-Free) antibody was purified by affinity chromatography.
Concentration:
1 mg/ml
Storage & Handling:
The antibody solution should be stored undiluted at 4 °C. This LEAF™ solution contains no preservative; handle under aseptic conditions.
Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For immunofluorescent staining, the suggested use of this reagent is ≤ 0.25 µg per 106 cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.
Application Notes:
Additional reported applications (for the relevant formats) include: immunoprecipitation1, in vitro and in vivo modulation of B cell responses2-4, and immunohistochemistry5,6 of acetone-fixed frozen sections and formalin-fixed paraffin-embedded sections. The LEAF™ purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm sterile-filtered) is recommended for functional assays (Cat. No. 103216).
Application References:
1. Coffman, R.L. 1982. Immunol. Rev. 69:5. 2. George, A., et al., 1994. J. Immunol. 152:1014. 3. Asensi, V., et al., 1989. Immunology 68:204. 4. Domiati-Saad, R., et al., 1993. J. Immunol. 151:5936. 5. Hata, H., et al., 2004. J. Clin. Invest. 114:582. 6. Monteith, C. E. et al., 1996. Can. J. Vet.Res. 60: 193. 7. Shih, F. F., et al., 2006. J. Immunol. 176:3438. 8. Chang, C. L-T., et al., 2007. J. Immunol. 178:6984. 9. Fazilleau, N., et al., 2007. Nature Immunol. 8:753. 10. Charles, N., et al., 2010. Nat. Med. 16:701. (FC) PubMed
BALB/c mouse splenocytes stained with LEAF™ purified RA3-6B2, followed by anti-rat IgG FITC
Description:
CD45R is an isoform of CD45, also known as B220. It is a member of the protein tyrosine phosphatase (PTP) family with a molecular weight 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, 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, I.S., 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.
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