The 22E5.5 monoclonal antibody specifically binds to the extracellular domain of mouse Notch1. This Type 1 transmembrane protein is a member of the Notch family that includes Notch1-4. Notch ligands include membrane-bound Jagged1, Jagged2 and Delta-like1, 3 and 4. Notch receptors mediate juxtacrine signals between adjacent cells. Upon ligand binding, Notch1 undergoes proteolytic cleavage that results in the release of the Notch intracellular domain, NICD. NICD translocates to the nucleus where it forms a transcriptional activator complex with the RBP-J DNA binding factor and other transcriptional cofactors. These multimeric complexes regulate the expression of multiple genes including those that orchestrate many facets of embryonic development and the subsequent functioning of multiple organ systems such as the immune and nervous systems. Within the immune system, Notch signaling significantly affects the differentiation and survival of numerous cell types including thymocytes and subsets of T and B lymphocytes and dendritic cells. In altered forms, Notch1 has been implicated in transformation or progression in some T-cell neoplasms. The mN1A monoclonal antibody specifically binds to an intracellular domain of mouse Notch1.
The antibody was conjugated to BD Horizon™ BV650 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 650-nm. BD Horizon BV650 can be excited by the violet laser and detected in a filter used to detect APC-like dyes (eg, 660/20-nm filter). Due to the excitation and emission characteristics of the acceptor dye, there will be spillover into the APC and Alexa Fluor® 700 detectors. However, the spillover can be corrected through compensation as with any other dye combination.