RY586 Mouse Anti-Human CD45RO

BD Horizon™ RY586 Mouse Anti-Human CD45RO

Name: RY586 Mouse Anti-Human CD45RO
Brand: BD Horizon™
SKU: 568474

Clone UCHL1

(RUO)

Multiparameter flow cytometric analysis of CD45RO expression on Human peripheral blood leucocyte populations. Human whole blood was stained with APC Mouse Anti-Human CD45RA antibody (Cat. No. 550855/561884) and with either BD Horizon™ RY586 Mouse IgG2a, κ Isotype Control (Cat. No. 568131; Left Plots) or BD Horizon™ RY586 Mouse Anti-Human CD45RO antibody (Cat. No. 568473/568474; Right Plots). Erythrocytes were lysed with BD FACS™ Lysing Buffer (Cat. No. 349202). Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Upper 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 leucocyte populations. Bottom Plots: The bivariate pseudocolor density plot showing CD45RO expression (or Ig isotype control staining) versus CD45RA was derived from gated events with the side and forward light-scatter characteristics of intact lymphocytes.

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Product Details

Brand: BD Horizon™

Alternative Name: CD45R; PTPRC; LCA; Leukocyte common antigen; GP180; LY5; T200

Reactivity: Human (QC Testing)

Isotype: Mouse BALB/c IgG2a, κ

Immunogen: Human IL-2-dependent T-cell line

Application: Flow cytometry (Routinely Tested)

Vol. Per Test: 5 µl

Workshop Number: IV N31

Entrez Gene ID: 5788

Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.

Regulatory Status: RUO

Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. 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.

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.
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).
An isotype control should be used at the same concentration as the antibody of interest.
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.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
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).
CF™ is a trademark of Biotium, Inc.

Companion Products

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

RY586 Mouse IgG2a, κ Isotype Control RUO

Size 50 µg Cat No. 568131

APC Mouse Anti-Human CD45RA RUO

Size 100 Tests Cat No. 550855

Lysing Solution 10X Concentrate IVD

Size 100 mL Cat No. 349202

Lysing Buffer RUO

Size 100 mL Cat No. 555899

View Details

568474 Rev. 2

Antibody Details

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.

568474 Rev. 2

Format Details

RY586

The BD Horizon RealYellow™ 586 (RY586) Dye is part of the BD family of yellow-green dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 565-nm and an emission maximum (Em Max) at 586-nm. Driven by BD innovation, RY586 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), RY586 can be used as an alternative to PE and we recommend using an optical filter centered near 586-nm (eg, a 586/15-nm bandpass filter). For spectral instruments equipped with a Yellow-Green laser (561-nm), it can be used in conjunction with PE. Compared to PE, RY586 is similar in brightness, minimal spillover into Blue detectors, and increased spillover into the 610/20-nm (PE-CF594) detector. Please ensure that your instrument configuration (lasers and optical filters) is appropriate for this dye.

Format: RY586

Excitation Source: Yellow-Green 561 nm

Excitation Max: 564 nm

Emission Max: 586 nm

568474 Rev.2

Citations & References

Development References (6)

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
Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
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
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
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
Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.

View All (6)

568474 Rev. 2

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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

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.