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RY610 Mouse Anti-Human CD45RO
RY610 Mouse Anti-Human CD45RO
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.
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.
Product Details
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BD Horizon™
CD45R; PTPRC; LCA; Leukocyte common antigen; GP180; LY5; T200
Human (QC Testing)
Mouse BALB/c IgG2a, κ
Human IL-2-dependent T-cell line
Flow cytometry (Routinely Tested)
5 µl/test
IV N31
5788
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

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

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  3. For U.S. patents that may apply, see bd.com/patents.
  4. 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.
  5. 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).
  6. 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.
  7. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. An isotype control should be used at the same concentration as the antibody of interest.
  10. CF™ is a trademark of Biotium, Inc.
571355 Rev. 1
Antibody Details
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UCHL1

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.

571355 Rev. 1
Format Details
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RY610
The BD Horizon RealYellow™ 610 (RY610) Dye is part of the BD® family of yellow-green dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 557-nm and an emission maximum (Em Max) at 610-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RY610 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY610 can be used as an alternative to PE-CF594 and we recommend using an optical filter centered near 610-nm (eg, a 610/20-nm bandpass filter). For spectral instruments equipped with a Yellow-Green laser (561-nm), it can be used in conjunction with PE-CF594.
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RY610
Yellow-Green 561 nm
557 nm
610 nm
571355 Rev.1
Citations & References
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View product citations for antibody "571355" on CiteAb

Development References (6)

  1. 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
  2. Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
  3. 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
  4. 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
  5. 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
  6. Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
View All (6) View Less
571355 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.