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Multiparameter flow cytometric analysis using BD OptiBuild™ BV421 Mouse Anti-Human CD277 antibody (Cat. No. 742826) on human peripheral blood. Flow cytometry was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.
BD OptiBuild™ BV421 Mouse Anti-Human CD277
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
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.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- 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.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- 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.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
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Companion Products
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.
Development References (6)
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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
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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
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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
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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
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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
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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
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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.