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BUV661 Mouse Anti-Human HLA-G
Product Details
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BD OptiBuild™
HLA-G; HLAG; MHC class I antigen G; MHC-G; sHLA-G
Human (Tested in Development)
Mouse IgG2a, κ
HLA-G-transfected Cells
Flow cytometry (Qualified)
0.2 mg/ml
AB_2875651
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 CompBead 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 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. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

Note:  When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed.  For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).

Product Notices

  1. 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.
  2. Researchers should determine the optimal concentration of this reagent for their individual applications.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  7. 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.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. BD Horizon Brilliant Ultraviolet 661 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
751664 Rev. 1
Antibody Details
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87G

The 87G monoclonal antibody specifically recognizes Human Leukocyte Antigen G (HLA-G) which is encoded by HLA-G (major histocompatibility complex, class I, G). HLA-G is a nonclassical Major Histocompatibility Complex class I (MHC-Ib) molecule that is structurally related to the classical MHC class Ia antigens (HLA-A, -B, -C). Several HLA-G isoforms have been described including transmembrane HLA-G1, -G2, -G3, -G4 and soluble HLA-G5, -G6, and -G7. The 87G monoclonal antibody reportedly recognizes a conformationally-dependent epitope on the heterodimeric transmembrane HLA-G1 and soluble HLA-G7 isoforms that consist of an HLA-G alpha chain and β2-microglobulin (β2m). HLA-G1 is variably expressed on placental trophoblast cells, thymic epithelial cells, activated monocytes, macrophages, dendritic cells, and tumor cells. Heterodimeric HLA-G shows limited variation and binds a limited variety of self-peptides derived from intracellular proteins including histones and ribosomal proteins. This molecule binds to inhibitory receptors such as CD85d, CD85j, and CD158d that are differentially expressed by NK cells, T cells, monocytes, dendritic cells, and B cells. This interaction exerts suppressive regulation of immune responses and is thought to help safeguard maternal tolerance of the fetus during pregnancy. The 4H84 monoclonal antibody that reportedly recognizes denatured forms of HLA-G1 and HLA-G2 has also been described.

The antibody was conjugated to BD Horizon™ BUV661 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 with an Ex Max of 348-nm and an acceptor dye with an Em Max at 661-nm. BD Horizon Brilliant BUV661 can be excited by the ultraviolet laser (355 nm) and detected with a 670/25 filter and a 630 nm LP.  Due to cross laser excitation of this dye, there may be significant spillover into channels detecting APC-like emissions (eg, 670/25-nm filter).

    

Due to spectral differences between labeled cells and beads, using BD™ CompBeads can result in incorrect spillover values when used with BD Horizon BUV661 reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different BUV661 reagents (eg, CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone-specific compensation controls when using these reagents.

751664 Rev. 1
Format Details
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BUV661
The BD Horizon Brilliant™ Ultraviolet 661 (BUV661) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This tandem fluorochrome is comprised of a BUV395 donor with an excitation maximum (Ex Max) of 350-nm and an acceptor dye with an emission maximum (Em Max) at 660-nm. BUV661, driven by BD innovation, is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 660-nm (e.g., 670/25 bandpass filter). The acceptor dye can be excited by the Red (628–640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BUV661
Ultraviolet 355 nm
350 nm
660 nm
751664 Rev.1
Citations & References
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Development References (6)

  1. Dorling A, Monk NJ, Lechler RI. HLA-G inhibits the transendothelial migration of human NK cells. Eur J Immunol. 2000; 30(2):586-593. (Biology). View Reference
  2. King A, Allan DS, Bowen M, et al. HLA-E is expressed on trophoblast and interacts with CD94/NKG2 receptors on decidual NK cells. Eur J Immunol. 2000; 30(6):1623-1631. (Biology). View Reference
  3. Le Gal FA, Riteau B, Sedlik C, et al. HLA-G-mediated inhibition of antigen-specific cytotoxic T lymphocytes. Int Immunol. 1999; 11(8):1351-1356. (Biology). View Reference
  4. Lee N, Malacko AR, Ishitani A, et al. The membrane-bound and soluble forms of HLA-G bind identical sets of endogenous peptides but differ with respect to TAP association. Immunity. 1995; 3(5):591-600. (Immunogen: ELISA, Flow cytometry). View Reference
  5. Odum N, Ledbetter JA, Martin P, et al. Homotypic aggregation of human cell lines by HLA class II-, class Ia- and HLA-G-specific monoclonal antibodies.. Eur J Immunol. 1991; 21(9):2121-31. (Immunogen: Functional assay). View Reference
  6. Yang Y, Chu W, Geraghty DE, Hunt JS. Expression of HLA-G in human mononuclear phagocytes and selective induction by IFN-gamma.. J Immunol. 1996; 156(11):4224-31. (Clone-specific: Flow cytometry, Immunohistochemistry). View Reference
View All (6) View Less
751664 Rev. 1

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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.