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RB670 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.

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

  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. For U.S. patents that may apply, see bd.com/patents.
  4. 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.
  5. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  6. 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.
  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. An isotype control should be used at the same concentration as the antibody of interest.
  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.
  13. 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.
771599 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.

771599 Rev. 1
Format Details
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RB670
The BD Horizon RealBlue™ 670 (RB670) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 492 nm and an emission maximum (Em Max) at 670 nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB670 can be used on both spectral and conventional cytometers and is designed to be primarily excited by the Blue laser (488-nm). For conventional instruments equipped with only a Blue laser (488-nm), RB670 can be used as an alternative to PE-Cy5 and we recommend using an optical filter centered near 670-nm (eg, a 670/30-nm bandpass filter). For conventional and spectral instruments equipped with both a Blue (488-nm) and Yellow-Green (561-nm) laser and appropriate detectors, it can be used in conjunction with PE-Cy5.
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RB670
Blue 488 nm
492 nm
670 nm
771599 Rev.1
Citations & References
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View product citations for antibody "771599" on CiteAb

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
771599 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.