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 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
- 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).
- CF™ is a trademark of Biotium, Inc.
- BD Horizon Brilliant Ultraviolet 615 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.
- Although hamster immunoglobulin isotypes have not been well defined, BD Biosciences Pharmingen has grouped Armenian and Syrian hamster IgG monoclonal antibodies according to their reactivity with a panel of mouse anti-hamster IgG mAbs. A table of the hamster IgG groups, Reactivity of Mouse Anti-Hamster Ig mAbs, may be viewed at http://www.bdbiosciences.com/documents/hamster_chart_11x17.pdf.
Companion Products
The 2C9.G2 monoclonal antibody specifically binds to the integrin β3 chain (CD61), which associates with the integrin αv chain (CD51) to form the vitronectin receptor, as well as the αIIb chain (CD41) to form the gpIIb/IIIa complex. Both receptors mediate adhesion to fibronectin, fibrinogen, vitronectin, thrombospondin, and von Willebrand factor. Leukocyte-endothelial adhesion is also mediated by the binding of αvβ3 integrin or vitronectin receptor to CD31 (PECAM-1). In addition, interaction of the αvβ3 integrin with its ligands regulates the L-type Ca2+ channel in vascular smooth muscle cells, possibly mediating vasodilatory responses to injury. Soluble and insoluble 2C9.G2 mAb mimics the effect of the natural ligands in smooth muscle cells from rat cremaster arterioles. Furthermore, osteopontin, also named Eta-1, is a cytokine that binds to αvβ3. CD61 is expressed on platelets, activated T lymphocytes, polymorphonuclear granulocytes, and blastocysts. Cross-reactivity of mAb 2C9.G2 to rat mast cells and platelets has been observed by flow cytometric analysis. mAb 2C9.G2 has been demonstrated to block binding of rat and mouse cells to fibronectin.
The antibody was conjugated to BD Horizon BUV615 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome with an Ex Max near 350 nm and an Em Max near 615 nm. BD Horizon Brilliant BUV615 can be excited by the ultraviolet laser (355 nm) and detected with a 610/20 filter and a 595 nm LP. Due to the excitation of the acceptor dye by the blue/yellow-green laser line, there may be significant spillover into channels detecting PE-CF594 like emissions (eg, 610/20-nm filter).
Development References (8)
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Ashkar S, Weber GF, Panoutsakopoulou V, et al. Eta-1 (osteopontin): an early component of type-1 (cell-mediated) immunity. Science. 2000; 287(5454):860-864. (Clone-specific: Blocking). View Reference
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Frieser M, Hallmann R, Johansson S, Vestweber D, Goodman SL, Sorokin L. Mouse polymorphonuclear granulocyte binding to extracellular matrix molecules involves beta 1 integrins. Eur J Immunol. 1996; 26(12):3127-3136. (Biology). View Reference
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Kieffer N, Phillips DR. Platelet membrane glycoproteins: functions in cellular interactions. Annu Rev Cell Biol. 1990; 6:329-357. (Biology). View Reference
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Moulder K, Roberts K, Shevach EM, Coligan JE. The mouse vitronectin receptor is a T cell activation antigen. J Exp Med. 1991; 173(2):343-347. (Biology). View Reference
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Piali L, Hammel P, Uherek C, et al. CD31/PECAM-1 is a ligand for alpha v beta 3 integrin involved in adhesion of leukocytes to endothelium. J Cell Biol. 1995; 130(2):451-460. (Clone-specific: Blocking). View Reference
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Schultz JF, Armant DR. Beta 1- and beta 3-class integrins mediate fibronectin binding activity at the surface of developing mouse peri-implantation blastocysts. Regulation by ligand-induced mobilization of stored receptor. J Biol Chem. 1995; 270(19):11522-11531. (Clone-specific: Blocking). View Reference
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Wu X, Mogford JE, Platts SH, Davis GE, Meininger GA, Davis MJ . Modulation of calcium current in arteriolar smooth muscle by alphav beta3 and alpha5 beta1 integrin ligands. J Cell Biol. 1998; 143(1):241-252. (Biology). View Reference
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Yasuda M, Hasunuma Y, Adachi H, et al. Expression and function of fibronectin binding integrins on rat mast cells.. Int Immunol. 1995; 7(2):251-8. (Immunogen: Flow cytometry, Functional assay, Immunofluorescence, Immunoprecipitation, Inhibition). 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.