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This antibody has been reported to react with the 25kDa ε chain of the T-cell receptor-associated CD3 complex expressed on thymocytes, mature T lymphocytes, and NK-T cells on many mouse stains. Plate-bound and soluble hamster anti-mouse CD3e (clone 500A2) antibody has been reported to activate unprimed T cells in vitro. Activation of a mouse T-cell clone by soluble hamster anti-mouse CD3e (clone 500A2) can be blocked by Fab fragments of anti-CD4 antibody GK1.5, suggesting that the hamster anti-mouse CD3e (clone 500A2) antibody may bind an epitope of CD3e close to a site at which CD4 associates with the T-cell receptor. This hamster mAb to a mouse leukocyte antigen has been reported not to cross-react with rat leukocytes. 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 415 nm 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 beads in 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.Flow Cytometry