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Multiparameter flow cytometric analysis of CD45RO expression on Human peripheral blood leukocyte populations. Human whole blood was stained with BD Horizon™ BV421 Mouse Anti-Human CD45RA antibody (Cat. No. 562885; Bottom Plots) and with either BD Horizon™ RY610 Mouse IgG2a, κ Isotype Control (Cat. No. 571157; Left Plots) or BD Horizon™ RY610 Mouse Anti-Human CD45RO antibody (Cat. No. 571351/571355; Right Plots). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer and FlowJo™ Software. Top Plots: The bivariate pseudocolor density plot showing CD45RO expression (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the side and forward light-scatter characteristics of intact leukocytes. Bottom Plots: The bivariate pseudocolor density plot showing CD45RO expression (or Ig Isotype control staining) versus CD45RA was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes.
BD Horizon™ RY610 Mouse Anti-Human CD45RO
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Preparation And Storage
Recommended Assay Procedures
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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The UCHL1 monoclonal antibody specifically binds to the 180 kDa isoform of CD45 (aka, the Leukocyte Common Antigen). CD45RO is a type I transmembrane glycoprotein that has cytoplasmic protein tyrosine phosphatase activity and functions in signal transduction pathways. This CD45 isoform does not include amino acid sequences encoded by the variable CD45 exons A, B, or C. CD45RO is expressed on most thymocytes, activated T cells, memory T cells, granulocytes and monocytes, but only on a proportion of resting T cells. CD45RO and CD45RA antibodies seem to define complementary, predominantly non-overlapping, populations in resting peripheral T cells, demonstrating heterogeneity within the CD8 and CD4 subpopulations. CD45RO binds to CD22.
Development References (6)
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Akbar AN, Terry L, Timms A, Beverley PC, Janossy G. Loss of CD45R and gain of UCHL1 reactivity is a feature of primed T cells. J Immunol. 1988; 140(7):2171-2178. (Clone-specific: Flow cytometry). View Reference
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Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
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Norton AJ, Ramsay AD, Smith SH, Beverley PC, Isaacson PG. Monoclonal antibody (UCHL1) that recognises normal and neoplastic T cells in routinely fixed tissues. J Clin Pathol. 1986; 39(4):399-405. (Immunogen: Immunohistochemistry). View Reference
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Smith SH, Brown MH, Rowe D, Callard RE, Beverley PC. Functional subsets of human helper-inducer cells defined by a new monoclonal antibody, UCHL1. Immunology. 1986; 58(1):63-70. (Clone-specific: Flow cytometry, Immunohistochemistry, Immunoprecipitation). View Reference
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Streuli M, Morimoto C, Schrieber M, Schlossman SF, Saito H. Characterization of CD45 and CD45R monoclonal antibodies using transfected mouse cell lines that express individual human leukocyte common antigens. J Immunol. 1988; 141(11):3910-3914. (Clone-specific: Flow cytometry). View Reference
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Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
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