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      RB705 Mouse Anti-Human CD86 (B7-2)
      RB705 Mouse Anti-Human CD86 (B7-2)

      Multiparameter flow cytometric analysis of CD86 (B7-2) expression on Human peripheral blood leukocyte populations. Fresh whole blood was first treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. After washing, the leukocytes were preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219) and then stained with either BD Horizon™ RB705 Mouse IgG2b, κ Isotype Control (Cat. No. 570273; Left Plot) or BD Horizon™ RB705 Mouse Anti-Human CD86 (B7-2) antibody (Cat. No. 570615/570703; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD86 (B7-2) [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 BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.

      RB705 Mouse Anti-Human CD86 (B7-2)

      Multiparameter flow cytometric analysis of CD86 (B7-2) expression on Human peripheral blood leukocyte populations. Fresh whole blood was first treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. After washing, the leukocytes were preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219) and then stained with either BD Horizon™ RB705 Mouse IgG2b, κ Isotype Control (Cat. No. 570273; Left Plot) or BD Horizon™ RB705 Mouse Anti-Human CD86 (B7-2) antibody (Cat. No. 570615/570703; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD86 (B7-2) [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 BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.

      Product Details
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      BD Horizon™
      B7.2; B7-2; B-lymphocyte activation antigen B7-2; B70; BU63; CD28LG2; LAB72
      Human (QC Testing), Rhesus,Cynomolgus,Baboon (Reported)
      Mouse IgG2b, κ
      Human B-Lymphoblastoid Cell Line (JY)
      Flow cytometry (Routinely Tested)
      5 µl/test
      VI CD86.8
      942
      AB_3685894
      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.
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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. 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.
      2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      3. 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).
      4. An isotype control should be used at the same concentration as the antibody of interest.
      5. 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.
      6. 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.
      7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      8. 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.
      9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      10. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
      11. For U.S. patents that may apply, see bd.com/patents.
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      570703 Rev. 1
      Antibody Details
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      IT2.2

      The IT2.2 monoclonal antibody specifically binds to CD86. CD86 is a ~ 75 kDa type I transmembrane glycoprotein which is also known as B7-2 or B70. CD86 is primarily expressed on monocytes, dendritic cells, Langerhans cells, and activated B cells including B lymphoid cells in germinal centers and Epstein-Barr virus transformed B-cell lines. CD86 is a ligand for CD28 and CD152 (CTLA-4) and may play an important role in costimulation of T cells in primary immune responses. Competitive binding assays demonstrate that, while both the IT2.2 and FUN-1 monoclonal antibodies recognize the same CD86 molecule, they react with different epitopes. IT2.2 blocks the costimulation activity of CD86 in functional studies and blocks binding of human CTLA-4-Ig fusion protein to CD86 gene-transfected cells.

      570703 Rev. 1
      Format Details
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      RB705
      The BD Horizon RealBlue™ 705 (RB705) 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 707-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB705 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), RB705 can be used as an alternative to PerCP-Cy5.5 or BB700 and we recommend using an optical filter centered near 710-nm (e.g., a 695/40 or 710/50-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PerCP-Cy5.5. RB705 is on average brighter than PerCP-Cy5.5 and BB700, and has minimal spillover into Yellow-Green detectors.
      altImg
      RB705
      Blue 488 nm
      498 nm
      707 nm
      570703 Rev.1
      Citations & References
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      View product citations for antibody "570703" on CiteAb

      Development References (7)

      1. Azuma M, Ito D, Yagita H, et al. B70 antigen is a second ligand for CTLA-4 and CD28.. Nature. 1993; 366(6450):76-9. (Immunogen: Blocking, Flow cytometry). View Reference
      2. Bjornson-Hooper ZB, Fragiadakis GK, Spitzer MH, et al. A Comprehensive Atlas of Immunological Differences Between Humans, Mice, and Non-Human Primates.. Front Immunol. 2022; 13:867015. (Clone-specific: Flow cytometry). View Reference
      3. Engel P, Gribben JG, Freeman GJ, et al. The B7-2 (B70) costimulatory molecule expressed by monocytes and activated B lymphocytes is the CD86 differentiation antigen. Blood. 1994; 84(5):1402-1407. (Biology). View Reference
      4. Engel P, Wagner N, Tedder TF. CD86 Workshop Report. In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:703-705.
      5. Hardie DL, Casamayor M, Johnson GD, et al. CD86 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.; 1997:201-204.
      6. Nozawa Y, Wachi E, Tominaga K, Abe M, Wakasa H. A novel monoclonal antibody (FUN-1) identifies an activation antigen in cells of the B-cell lineage and Reed-Sternberg cells. J Pathol. 1993; 169(3):309-315. (Biology). View Reference
      7. Waggie KS, Holdren MS, Byrnes-Blake K, et al. Preclinical safety, pharmacokinetics, and pharmacodynamics of recombinant human interleukin-21 in cynomolgus macaques (Macaca fascicularis).. Int J Toxicol. 2012; 31(4):303-16. (Clone-specific: Flow cytometry). View Reference
      View All (7) View Less
      570703 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.