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Flow cytometric analysis of CD45 expression on rat splenocytes. Lewis rat splenic leucocytes were preincubated with Purified Mouse Anti-Rat CD32 antibody (Rat BD Fc Block™) (Cat. No. 550270/550271). The cells were then stained with either BD Horizon™ R718 Mouse Anti-Rat CD45 antibody (Cat. No. 567734; solid line histogram) or BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928; dashed line histogram) at 1.0 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histograms showing CD45 expression (or Ig Isotype control staining) were derived from events with the forward and side light-scatter characteristics of viable (DAPI-negative) splenocytes. Flow cytometry and data analysis was performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Horizon™ R718 Mouse Anti-Rat CD45
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BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
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- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
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The OX-1 antibody recognizes all molecular forms of CD45 (Leukocyte Common Antigen) on all hematopoietic cells except erythrocytes. CD45 is a member of the Protein Tyrosine Phosphatase (PTP) family; its intracellular (COOH-terminal) region contains two PTP catalytic domains while the extracellular region is highly variable due to alternative splicing of exons 4, 5, and 6 (designated A, B, and C, respectively), and differing levels of glycosylation. The CD45 isoforms detected in rat are cell type-, maturation-, and activation state-specific. The CD45 isoforms play complex roles in T-cell and B-cell antigen receptor signal transduction.
The antibody was conjugated to BD Horizon™ Red 718, which has been developed exclusively by for BD Biosciences as a better alternative to Alexa Fluor™ 700. BD Horizon™ Red 718 can be excited by the red laser (628 – 640 nm) and, with an Em Max around 718 nm, it can be detected using a 730/45 nm filter. Due to similar excitation and emission properties, we do not recommend using R718 in combination with APC-R700 or Alexa Fluor™ 700.
研发参考 (4)
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Harb, R., Xie, et al. Bone marrow progenitor cells repair rat hepatic sinusoidal endothelial cells after liver injury. Gastroenterology.. 2009; 137(2):704-12. (Clone-specific: Flow cytometry, Immunocytochemistry, Immunofluorescence, Immunohistochemistry). 查看参考
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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.
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Sunderland CA, McMaster WR, Williams AF. Purification with monoclonal antibody of a predominant leukocyte-common antigen and glycoprotein from rat thymocytes. Eur J Immunol. 1979; 9(2):155-159. (Immunogen: Flow cytometry). 查看参考
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Woollett GR, Barclay AN, et al. Molecular and antigenic heterogeneity of the rat leukocyte common antigen from thymocytes and T and B lymphocytes. . Eur J Immunol. 1985; 15:168-173. (Biology).
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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.
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