The MHN1-519 monoclonal antibody specifically binds to an extracellular domain of human Notch1. Notch1 is a type 1 transmembrane glycoprotein receptor and member of the Notch family that includes Notch1-Notch4. Notch1 is cleaved in the Golgi and presents as a cell surface heterodimeric receptor. The Notch1 receptor can bind to several membrane-bound ligands including Jagged1, Jagged2, Delta1 and Delta4. 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 various transcriptional factors. These multimeric complexes either positively or negatively 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, cardiovascular and nervous systems. Within the immune system, Notch signaling significantly affects the development, proliferation, 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 associated with certain cardiovascular diseases and with some lymphocyte neoplasms.
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.