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      RY586 Mouse Anti-Human CD277
      Product Details
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      BD OptiBuild™
      CD277; Butyrophilin 3; BTN3; BTN3A1, BTN3A2, BTN3A3; BTN3.1, BTN3.2, BTN3.3
      Human (Tested in Development)
      Mouse BALB/c IgG2a, κ
      Human BTN3A3
      Flow cytometry (Qualified)
      0.2 mg/ml
      10384
      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 that minimize unconjugated dye and antibody.
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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. Researchers should determine the optimal concentration of this reagent for their individual applications.
      2. 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.
      3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      4. An isotype control should be used at the same concentration as the antibody of interest.
      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. CF™ is a trademark of Biotium, Inc.
      7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      9. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      10. 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.
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      753570 Rev. 1
      Antibody Details
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      232-5

      The 232-5 monoclonal antibody specifically recognizes Butyrophilin 3 (BTN3), which is also known as CD277. BTN3 is comprised of three structurally related members, BTN3A1, BTN3A2, and BTN3A3. BTN3 molecules are transmembrane type 1 glycoproteins that belong to the Ig superfamily. The 232-5 antibody recognizes the variable region of the extracellular domains of BTN3 molecules. BTN3 molecules are differentially expressed by T, NKT, B, and NK cells, monocytes, macrophages, dendritic cells, as well as some stem cells and tumor cells. BTN3 expression is decreased upon lymphocyte activation. The extracellular domain of CD277 is structurally related to the B7 family of proteins that regulate T cell activation. A soluble form of the 232-5 antibody can serve as an agonist in BTN3-mediated regulation of anti-CD3 ± anti-CD28 antibody activated CD4+ or CD8+ T cells.

      753570 Rev. 1
      Format Details
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      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.
      altImg
      RY586
      Yellow-Green 561 nm
      564 nm
      586 nm
      753570 Rev.1
      Citations & References
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      Development References (6)

      1. Afrache H, Gouret P, Ainouche S, Pontarotti P, Olive D. The butyrophilin (BTN) gene family: from milk fat to the regulation of the immune response.. Immunogenetics. 2012; 64(11):781-94. (Clone-specific: Functional assay). View Reference
      2. Cubillos-Ruiz JR, Martinez D, Scarlett UK, et al. CD277 is a negative co-stimulatory molecule universally expressed by ovarian cancer microenvironmental cells.. Oncotarget. 2010; 1(5):329-38. (Biology). View Reference
      3. Harly C, Guillaume Y, Nedellec S, et al. Key implication of CD277/butyrophilin-3 (BTN3A) in cellular stress sensing by a major human γδ T-cell subset.. Blood. 2012; 120(11):2269-79. (Biology). View Reference
      4. Kabelitz D. Critical role of butyrophilin 3A1 in presenting prenyl pyrophosphate antigens to human γδ T cells.. Cell Mol Immunol. 2014; 11(2):117-9. (Clone-specific: Functional assay). View Reference
      5. Messal N, Mamessier E, Sylvain A, et al. Differential role for CD277 as a co-regulator of the immune signal in T and NK cells.. Eur J Immunol. 2011; 41(12):3443-54. (Biology). View Reference
      6. Yamashiro H, Yoshizaki S, Tadaki T, Egawa K, Seo N. Stimulation of human butyrophilin 3 molecules results in negative regulation of cellular immunity.. J Leukoc Biol. 2010; 88(4):757-67. (Immunogen: Blocking, Flow cytometry, Functional assay, Immunoprecipitation, Inhibition, Stimulation, Western blot). View Reference
      View All (6) View Less
      753570 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.

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