The B81-3 monoclonal antibody was raised against keyhole limpet hemocyanin (KLH), an antigen not expressed by mammalian cells or cell lines. Intended for use as a hamster immunoglobulin isotype control, the B81-3 antibody can be used as a negative control to help differentiate non-specific background signal when compared against a signal from a specific hamster antibody. The purified B81-3 antibody may also serve as a useful standard for some ELISA applications. KLH is a oxygen carrying metalloprotein from a species of keyhole limpet that lives off the coast of California and is often used as a carrier protein to help elicit immune responses to haptens. Since KLH is phylogenetically distant from many mammalian proteins, it often has very minimal to no cross-reactivity for many immunological assays.
The antibody was conjugated to BD Horizon BUV737 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 with an Ex Max of 348-nm and an acceptor dye with an Em Max at 737-nm. BD Horizon Brilliant BUV737 can be excited by the ultraviolet laser (355 nm) and detected with a 740/35 filter. Due to the excitation of the acceptor dye by other laser lines, there may be significant spillover into channels detecting Alexa Fluor® 700-like dyes (eg, 712/20-nm filter).
Due to spectral differences between labeled cells and beads, using BD™ CompBeads can result in incorrect spillover values when used with BD Horizon BUV737 reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different BUV737 reagents (eg, 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.
