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      RY775 Mouse Anti-Human CD27
      RY775 Mouse Anti-Human CD27
      Multiparameter flow cytometric analysis of CD27 expression on Human peripheral blood leukocyte populations. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leukocytes were washed, preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219) and stained with either BD Horizon™ RY775 Mouse IgG1, κ Isotype Control (Cat. No. 571408; Left Plot) or BD Horizon™ RY775 Mouse Anti-Human CD27 antibody (Cat. No. 571660/571738; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD27 (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 leukocytes. Flow cytometry and data analysis were performed using a 5-laser Flow Cytometer System and FlowJo™ Software.
      RY775 Mouse Anti-Human CD27
      Multiparameter flow cytometric analysis of CD27 expression on Human peripheral blood leukocyte populations. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leukocytes were washed, preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219) and stained with either BD Horizon™ RY775 Mouse IgG1, κ Isotype Control (Cat. No. 571408; Left Plot) or BD Horizon™ RY775 Mouse Anti-Human CD27 antibody (Cat. No. 571660/571738; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD27 (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 leukocytes. Flow cytometry and data analysis were performed using a 5-laser Flow Cytometer System and FlowJo™ Software.
      Product Details
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      BD Horizon™
      TNFRSF7; Tumor necrosis factor receptor superfamily, member 7; Tp55; S152
      Human (QC Testing)
      Mouse BALB/c IgG1
      Human Activated Peripheral Blood Cells
      Flow cytometry (Routinely Tested)
      5 µl/test
      VI T6T037
      939
      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 unconjugated antibody and free dye were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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      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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      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. An isotype control should be used at the same concentration as the antibody of interest.
      4. For U.S. patents that may apply, see bd.com/patents.
      5. 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.
      6. 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).
      7. 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.
      8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      9. 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.
      10. 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.
      11. Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
      12. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
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      571660 Rev. 1
      Antibody Details
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      L128

      The L128 monoclonal antibody specifically binds to human CD27. CD27 is a 55-kDa disulfide-linked dimer that is a member of the nerve growth factor (NGF) super family. This family also includes CD40, rat OX40, tumor necrosis factor (TNF) receptors and CD95 (Fas). With its ligand CD70, CD27 acts in a co-stimulatory fashion on T lymphocytes. Present on most peripheral blood T lymphocytes and medullary thymocytes, the CD27 antigen is upregulated upon activation with the release of a soluble form, 28 to 32 kDa. It is also detected on a subpopulation of approximately 33% of circulating B lymphocytes. Following exposure to antigens, CD45RA+ T lymphocytes respond by upregulating the CD27 antigen. After maximal stimulation, the CD27 antigen cannot be re-expressed on long-term cultures or on CD45RA-CD27+ T lymphocytes. The CD4+CD27- population is contained within the memory CD45RO+ subset that proliferates after exposure to allergens. Two subpopulations of B lymphocytes bearing the CD27 antigen secrete IgM (δ+) and IgG (δ-).

      571660 Rev. 1
      Format Details
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      RY775
      The BD Horizon RealYellow™ 775 (RY775) 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 775-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RY775 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), RY775 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).
      altImg
      RY775
      Yellow-Green 561 nm
      557 nm
      775 nm
      571660 Rev.1
      Citations & References
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      View product citations for antibody "571660" on CiteAb

      Development References (7)

      1. De Jong R, Brouwer M, Hooibrink B, Van der Pouw-Kraan T, Miedema F, Van Lier RA. The CD27- subset of peripheral blood memory CD4+ lymphocytes contains functionally differentiated T lymphocytes that develop by persistent antigenic stimulation in vivo. Eur J Immunol. 1992; 22(4):993-999. (Biology). View Reference
      2. Giesecke C, Meyer T, Durek P, et al. Simultaneous Presence of Non- and Highly Mutated Keyhole Limpet Hemocyanin (KLH)-Specific Plasmablasts Early after Primary KLH Immunization Suggests Cross-Reactive Memory B Cell Activation.. J Immunol. 2018; 200(12):3981-3992. (Clone-specific: Flow cytometry). View Reference
      3. Hintzen RQ, Lens SM, Beckmann MP, Goodwin RG, Lynch D, van Lier RA. Characterization of the human CD27 ligand, a novel member of the TNF gene family. J Immunol. 1994; 152(4):1762-1773. (Biology). View Reference
      4. Hintzen RQ, de Jong R, Lens SM, Brouwer M, Baars P, van Lier RA. Regulation of CD27 expression on subsets of mature T-lymphocytes. J Immunol. 1993; 151(5):2426-2435. (Biology). View Reference
      5. Kobata T, Agematsu K, Kameoka J, Schlossman SF, Morimoto C. CD27 is a signal-transducing molecule involved in CD45RA+ naive T cell costimulation. J Immunol. 1994; 153(12):5422-5432. (Biology). View Reference
      6. Kobata T, Morimoto C. CD27 Workshop Panel Report. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:67-69.
      7. Maurer D, Fischer GF, Fae I, et al. IgM and IgG but not cytokine secretion is restricted to the CD27+ B lymphocyte subset. J Immunol. 1992; 148(12):3700-3705. (Biology). View Reference
      View All (7) View Less
      571660 Rev. 1

      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.

      Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.