Multiparameter flow cytometric analysis using BD OptiBuild™ RB705 Mouse Anti-Human CD86 (B7-2) antibody (Cat. No. 757164) on Human peripheral blood. Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer.
Multiparameter flow cytometric analysis using BD OptiBuild™ RB705 Mouse Anti-Human CD86 (B7-2) antibody (Cat. No. 757164) on Human peripheral blood. Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer.
Product Details
BD OptiBuild™
B7.2; B7-2; B-lymphocyte activation antigen B7-2; B70; BU63; CD28LG2; LAB72
Human,Rhesus,Cynomolgus,Baboon (Tested in Development)
Mouse BALB/c IgG1, κ
Human HBL-1 Cell Line
Flow cytometry (Qualified)
0.2 mg/ml
V B046, BP126
942
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 that minimize unconjugated dye and antibody. 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
- 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.
- The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- 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.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- 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.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
Human BD Fc Block™ RUO
Size
0.25 mg
Cat No.
564220
RB705 Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
570261
757164 Rev. 2
Antibody Details
2331 (FUN-1)
The 2331 (FUN-1) monoclonal antibody specifically recognizes a 75 kDa transmembrane cell surface protein, CD86 (B70/B7-2), expressed primarily on monocytes, dendritic cells and activated B cells. Competitive binding assays demonstrate that, while both 2331 (FUN-1) and IT2.2 (Anti-CD86) antibodies specifically recognize the same molecule, they react with different epitopes. CD86 is a ligand for CD28 and CTLA-4 and plays an important role in costimulation of T cells in primary immune response. The 2331 (FUN-1) antibody blocks the costimulatory activity of CD86 when tested in functional studies.
757164 Rev. 2
Format Details
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.
RB705
Blue 488 nm
498 nm
707 nm
757164 Rev.2
Citations & References
View product citations for antibody "757164" on CiteAb
Development References (5)
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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. (Clone-specific: Blocking, Enhancement, Flow cytometry, Functional assay, Inhibition). View Reference
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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.
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Nozawa Y, Abe M, Wakasa H. Three mAb, FUN-1, FB1, and FB21, that recognize B-cell antigens in frozen or paraffin-embedded tissue sections. 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; 1995:705-706.
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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. (Immunogen: Flow cytometry, Fluorescence microscopy, Immunofluorescence, Immunohistochemistry, Immunoprecipitation). View Reference
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Nozawa Y, Wakasa H, Abe M. Production and usefulness of monoclonal antibodies against B cells. Fukushima J Med Sci. 1999; 45(1):1-11. (Clone-specific). View Reference
757164 Rev. 2
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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.