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 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 CompBead to ensure that BD Comp beads 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).
Product Notices
- 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.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- 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.
Companion Products
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™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.
Development References (6)
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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
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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
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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
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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
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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
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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
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.
