Preparation And Storage
Recommended Assay Procedures
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD OptiBuild Brilliant reagents) 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).
Product Notices
- This antibody was developed for use in flow cytometry.
- 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.
Companion Products
The GAP.A3 monoclonal antibody specifically recognizes the polymorphic HLA class I histocompatibility antigen, A-3 alpha chain (HLA-A3). This ~44 kDa human major histocompatibility complex (MHC) class I molecule noncovalently associates with the ~12 kDa monomorphic β2 microglobulin. Two major subtypes are encoded by HLA-A3, HLA-A3.1 and HLA-A3.2. HLA-A3 gene expression is more commonly detected in individuals from Europe and southern India. HLA-A3 is expressed on nearly all nucleated cells of HLA-A3-positive individuals. HLA-A3 functions in the presentation of antigens to CD8-positive T cells which may lead to the generation of HLA-A3-restricted cytotoxic T cells and memory T cells. When complexed with certain antigens, HLA-A3 can also bind to killer cell immunoglobulin-like receptors (KIR), such as KIR3DL2, and might regulate NK cell function.
Note: Since HLA-A3 expression varies between human populations, clone GAP.A3 staining can be donor-dependent. Based on in-house testing and current literature, individuals of European or Southern Indian descent more frequently express HLA-A3 than those of Asian descent. Data may differ between donors due to geographical variations of HLA-A3 expression.
The antibody was conjugated to BD Horizon™ BUV563 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 which has an Ex Max of 348 nm and an acceptor dye. The tandem has an Em Max at 563 nm. BD Horizon BUV563 can be excited by the 355 nm ultraviolet laser. On instruments with a 561 nm Yellow-Green laser, the recommended bandpass filter is 585/15 nm with a 535 nm long pass to minimize laser light leakage. When BD Horizon BUV563 is used with an instrument that does not have a 561 nm laser, a 560/40 nm filter with a 535 nm long pass may be more optimal. Due to the excitation and emission characteristics of the acceptor dye, there may be spillover into the PE and PE-CF594 detectors. However, the spillover can be corrected through compensation as with any other dye combination.
Development References (7)
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Allen TM, Altfeld M, Yu XG, et al. Selection, transmission, and reversion of an antigen-processing cytotoxic T-lymphocyte escape mutation in human immunodeficiency virus type 1 infection.. J Virol. 2004; 78(13):7069-78. (Biology). View Reference
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Augusto DG, O'Connor GM, Lobo-Alves SC, et al. Pemphigus is associated with KIR3DL2 expression levels and provides evidence that KIR3DL2 may bind HLA-A3 and A11 in vivo.. Eur J Immunol. 2015; 45(7):2052-60. (Biology). View Reference
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Berger AE, Davis JE, Cresswell P. Monoclonal antibody to HLA-A3.. Hybridoma. 1982; 1(2):87-90. (Immunogen). View Reference
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Carr TM, Adair SJ, Fink MJ, Hogan KT. Immunological profiling of a panel of human ovarian cancer cell lines.. Cancer Immunol Immunother. 2008; 57(1):31-42. (Clone-specific). View Reference
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Hansasuta P, Dong T, Thananchai H, et al. Recognition of HLA-A3 and HLA-A11 by KIR3DL2 is peptide-specific.. Eur J Immunol. 2004; 34(6):1673-9. (Biology). View Reference
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Lalouel JM, Jorde LB. Idiopathic hemochromatosis: significance and implications of linkage and association to HLA.. Ann N Y Acad Sci. 1988; 526:34-46. (Biology). View Reference
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Vonderheide RH, Anderson KS, Hahn WC, Butler MO, Schultze JL, Nadler LM. Characterization of HLA-A3-restricted cytotoxic T lymphocytes reactive against the widely expressed tumor antigen telomerase.. Clin Cancer Res. 2001; 7(11):3343-8. (Biology). 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.
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.