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The 3G8 monoclonal antibody specifically binds to the 50-65 kDa transmembrane form of the IgG Fc Receptor (FcγRIII), a human NK cell-associated antigen. CD16 is expressed on NK cells as well as macrophages and granulocytes. Reports indicate that CD16 plays a role in signal transduction and NK cell activation. The 3G8 antibody blocks the binding of soluble immune complexes to granulocytes. The 3G8 antibody is reported (Vossebeld et al., 1997) to increase intracellular calcium levels in human neutrophils by interacting with bothFcγRIIa and FcγRIIIb molecules. This antibody has also been reported to induce homotypic neutrophil aggregation. The antibody is conjugated to BD Horizon™ V500, which has been developed for use in multicolor flow cytometry experiments and is available exclusively from BD Biosciences. It is excited by the Violet laser with an Ex max of 415nm and Em Max at 500 nm. BD Horizon V500 conjugates emit at a similar wavelength to Amcyan yet exhibit reduced spillover into the FITC channel. For more information on BD Horizon V500, visit bdbiosciences.com/colors. When compensating dyes in this spectral range (such as Horizon™ V500 and AmCyan), the most accurate compensation can be obtained using single stained cellular controls. Due to spectral differences between cells and beadsin this channel, using BD CompBeads can result in spillover errors for V500 and AmCyan reagents.Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended.Different V500 reagents (e.g. 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.
