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.
- BD Horizon Brilliant™ Violet 750 is covered by one or more of the following US patents: 8,158,444; 8,802,450; 8,575,303; 8,455,613; 8,227,187; 8,841,072; 8,110,673.
Companion Products
The MBC 78.2 monoclonal antibody recognizes CD31 which is also known as, Platelet endothelial cell adhesion molecule (PECAM-1), platelet GPIIa, or EndoCAM. CD31 is a ~130 kDa type I transmembrane glycoprotein that belongs to the Ig gene superfamily. CD31 is comprised of an extracellular region with six IgC-like domains, a transmembrane region, and a cytoplasmic domain that contains two immunoreceptor tyrosine-based inhibitory motifs (ITIMs). The MBC 78.2 antibody specifically binds to an epitope located on membrane-proximal, extracellular Ig-like domain 6 of CD31. This epitope remains expressed by activated T cells after enzymatic cleavage and shedding of a soluble extracellular CD31 fragment comprised of Ig-like domains 1 to 5 from cells. In contrast to the MBC 78.2 antibody, the WM59 monoclonal antibody reportedly binds to the extracellular Ig-like domain 2 of CD31. WM59 can thus bind to cells that express intact CD31 but not to cells that express a truncated form CD31 that lacks at least the membrane distal Ig-like domains 1 and 2 of CD31. CD31 has wide tissue distribution and is expressed on platelets, monocytes, granulocytes, some T cell subsets, and at high levels on endothelial cells. This cell adhesion molecule has been implicated in a number of cellular phenomena, including vascular wound healing. angiogenesis, transendothelial migration of leucocytes, and the regulation of T cell responses.
The antibody was conjugated to BD Horizon™ BV750 which is part of the BD Horizon Brilliant™ Violet family of dyes. This dye is a tandem fluorochrome of BD Horizon BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 750-nm. BD Horizon Brilliant BV750 can be excited by the violet laser (405 nm) and detected with a 750/30 nm filter with a 740 nm long pass. Due to spectral differences between labeled cells and beads, using BD™ CompBeads can result in incorrect spillover values when used with BD Horizon BV750 reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended.
Development References (7)
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DeLisser HM, Chilkotowsky J, Yan HC, Daise ML, Buck CA, Albelda SM. Deletions in the cytoplasmic domain of platelet-endothelial cell adhesion molecule-1 (PECAM-1, CD31) result in changes in ligand binding properties.. J Cell Biol. 1994; 124(1-2):195-203. (Immunogen: Flow cytometry, Functional assay, Inhibition). View Reference
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Fawcett J, Buckley C, Holness CL, et al. Mapping the homotypic binding sites in CD31 and the role of CD31 adhesion in the formation of interendothelial cell contacts. J Cell Biol. 1995; 128(6):1229-1241. (Biology). View Reference
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Fornasa G, Groyer E, Clement M, et al. TCR stimulation drives cleavage and shedding of the ITIM receptor CD31. J Immunol. 2010; 184(10):5485-5492. (Clone-specific: Cytometric Bead Array, Flow cytometry, Immunofluorescence). View Reference
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Newman PJ, Paddock C. CD31 cluster workshop report. In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:1259-1260.
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Tanaka Y, Albelda SM, Horgan KJ, et al. CD31 expressed on distinctive T cell subsets is a preferential amplifier of beta 1 integrin-mediated adhesion.. J Exp Med. 1992; 176(1):245-53. (Clone-specific: Activation, Functional assay). View Reference
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Yan H-C, Newman PJ, Albelda SM. Epitope mapping of CD31 (PECAM-1) mAb. In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:1261-1262.
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Yan HC, Pilewski JM, Zhang Q, DeLisser HM, Romer L, Albelda SM. Localization of multiple functional domains on human PECAM-1 (CD31) by monoclonal antibody epitope mapping.. Cell Adhes Commun. 1995; 3(1):45-66. (Clone-specific: Immunoprecipitation). 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.
