The X40 hybridoma was derived from the fusion of mouse Sp2/0-Ag 14 myeloma cells with spleen cells from BALB/c mice immunized with keyhole limpet hemocyanin (KLH). The hybridoma secretes monoclonal antibody that binds specifically to KLH, an antigen not expressed on human cells or human cell lines.
The antibody was conjugated to BD Horizon™ BV711 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 711-nm. BD Horizon™ BV711 can be excited by the violet laser and detected in a filter used to detect Cy™5.5 / Alexa Fluor™ 700-like dyes (eg, 712/20-nm filter). Due to the excitation and emission characteristics of the acceptor dye, there may be moderate spillover into the Alexa Fluor™ 700 and PerCP-Cy5.5 detectors. However, the spillover can be corrected through compensation as with any other dye combination.