BD Horizon™ RB545 Mouse Anti-Human CD45RO
Clone UCHL1 (RUO)
Multiparameter flow cytometric analysis of CD45RO expression on Human peripheral blood leucocyte populations. Fresh whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. After washing, the leucocytes were preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219/564220) and then stained with BD Horizon™BV421 Mouse Anti-Human CD45RA antibody (Cat. No. 562885; Top Plots) and with either BD Horizon™ RB545 Mouse IgG2a, κ Isotype Control (Cat. No. 569277; Left Plots) or BD Horizon™ RB545 Mouse Anti-Human CD45RO antibody (Cat. No. 569750/569778; Right Plots). Samples were acquired using the BD FACSymphony™ A5 SE Cell Analyzer and spectrally unmixed using FlowJo™ v10.8 Software.
Top Plots: The bivariate pseudocolor density plot showing the correlated expression CD45RO (or Ig isotype control staining) versus CD45RA was derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes.
Bottom Plots: The bivariate pseudocolor density plot showing the correlated expression of CD45RO (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable leucocyte populations.
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Multiparameter flow cytometric analysis of CD45RO expression on Human peripheral blood leucocyte populations. Fresh whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. After washing, the leucocytes were preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219/564220) and then stained with BD Horizon™BV421 Mouse Anti-Human CD45RA antibody (Cat. No. 562885; Top Plots) and with either BD Horizon™ RB545 Mouse IgG2a, κ Isotype Control (Cat. No. 569277; Left Plots) or BD Horizon™ RB545 Mouse Anti-Human CD45RO antibody (Cat. No. 569750/569778; Right Plots). Samples were acquired using the BD FACSymphony™ A5 SE Cell Analyzer and spectrally unmixed using FlowJo™ v10.8 Software.
Top Plots: The bivariate pseudocolor density plot showing the correlated expression CD45RO (or Ig isotype control staining) versus CD45RA was derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes.
Bottom Plots: The bivariate pseudocolor density plot showing the correlated expression of CD45RO (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable leucocyte populations.
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.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- 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.
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
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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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For Research Use Only. Not for use in diagnostic or therapeutic procedures.