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BB700 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
AB_2743506
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 with BD Horizon BB700 under optimal conditions that minimize unconjugated dye and antibody.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794 or 566349).

When setting up compensation, it is recommended to compare spillover values obtained from cells and BD™ CompBeads to ensure that beads will provide sufficiently accurate spillover values.

For optimal results, it is recommended to perform two washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent staining prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

Product Notices

  1. This antibody was developed for use in flow cytometry.
  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. Researchers should determine the optimal concentration of this reagent for their individual applications.
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  8. 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.
  9. BD Horizon Brilliant Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
  10. Cy is a trademark of GE Healthcare.
746146 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.

The antibody was conjugated to BD Horizon™ BB700, which is part of the BD Horizon Brilliant™ Blue family of dyes.   It is a polymer-based tandem dye developed exclusively by BD Biosciences.  With an excitation max of 485 nm and an emission max of 693 nm, BD Horizon BB700 can be excited by the 488 nm laser and detected in a standard PerCP-Cy™5.5 set (eg, 695/40-nm filter). This dye provides a much brighter alternative to PerCP-Cy5.5 with less cross laser excitation off the 405 nm and 355 nm lasers.

746146 Rev. 1
Format Details
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BB700
The BD Horizon Brilliant™ Blue 700 (BB700) Dye is part of the BD Horizon Brilliant™ Blue family of dyes. This tandem fluorochrome is comprised of a polymer-technology dye donor with an excitation maximum (Ex Max) of 476-nm and an acceptor dye with an emission maximum (Em Max) at 695-nm. Driven by BD innovation, BB700 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 695-nm (e.g., a 695/20-nm bandpass filter). The donor dye can be excited by the Violet (405 nm) laser and the acceptor dye can be excited by the red (627–640 nm) laser resulting in cross-laser excitation and fluorescence spillover. BB700 Reagents are significantly brighter than equivalent PerCP or PerCP-Cy5.5 reagents and are less sensitive to photobleaching. In addition, BB700 shows much less excitation by the violet (407-nm) laser resulting in less spillover. BB700 has minimal yellow green (562-nm) excitation and is ideal for instruments with both blue (488-nm) and yellow green (562-nm) lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
BB700
Blue 488 nm
476 nm
695 nm
746146 Rev.1
Citations & References
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View product citations for antibody "746146" on CiteAb

Development References (3)

  1. 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. View Reference
  2. 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. View Reference
  3. 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. View Reference
746146 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.