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™ RB780 Mouse IgG2a, κ Isotype Control (Cat. No. 568740; Left Plots) or BD Horizon™ RB780 Mouse Anti-Human CD45RO antibody (Cat. No. 569124/569125; 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 the correlated expression of CD45RO (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 the correlated expression CD45RO (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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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™ RB780 Mouse IgG2a, κ Isotype Control (Cat. No. 568740; Left Plots) or BD Horizon™ RB780 Mouse Anti-Human CD45RO antibody (Cat. No. 569124/569125; 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 the correlated expression of CD45RO (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 the correlated expression CD45RO (or Ig isotype control staining) versus CD45RA was derived from gated events with the side and forward light-scatter characteristics of intact lymphocytes.
製品詳細
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
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.
推奨アッセイ手順
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
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- 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).
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
関連製品
Stain Buffer (FBS) RUO
サイズ
500 mL
カタログ番号
554656
Stain Buffer (BSA) RUO
サイズ
500 mL
カタログ番号
554657
RB780 Mouse IgG2a, κ Isotype Control RUO
サイズ
50 µg
カタログ番号
568740
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APC Mouse Anti-Human CD45RA RUO
サイズ
100 Tests
カタログ番号
550855
Lysing Solution 10X Concentrate RUO (GMP)
サイズ
100 mL
カタログ番号
349202
Lysing Buffer RUO
サイズ
100 mL
カタログ番号
555899
569124 Rev. 2
抗体の詳細
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.
569124 Rev. 2
フォーマットの詳細
RB780
The BD Horizon RealBlue™ 780 (RB780) Dye is part of the BD family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 781-nm. Driven by BD innovation, RB780 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB780 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-Cy7. RB780 is on average brighter than PE-Cy7 and has minimal spillover into Yellow-Green detectors.
RB780
Blue 488 nm
498 nm
781 nm
569124 Rev.2
引用＆参考文献
View product citations for antibody "569124" on CiteAb
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.
569124 Rev. 2
Please refer to Support Documents for Quality Certificates
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.