The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/100 - 1/500. Epitope exposure is recommended, with citrate buffer to enhance staining. Likely to work with frozen sections.
FunctionFunctions as a receptor for membrane-bound ligands Jagged1, Jagged2 and Delta1 to regulate cell-fate determination. Upon ligand activation through the released notch intracellular domain (NICD) it forms a transcriptional activator complex with RBPJ/RBPSUH and activates genes of the enhancer of split locus. Affects the implementation of differentiation, proliferation and apoptotic programs (By similarity). Involved in bone remodeling and homeostasis. In collaboration with RELA/p65 enhances NFATc1 promoter activity and positively regulates RANKL-induced osteoclast differentiation. Positively regulates self-renewal of liver cancer cells (PubMed:25985737).
Tissue specificityExpressed in the brain, heart, kidney, lung, skeletal muscle and liver. Ubiquitously expressed in the embryo.
Involvement in diseaseAlagille syndrome 2 Hajdu-Cheney syndrome
Sequence similaritiesBelongs to the NOTCH family. Contains 6 ANK repeats. Contains 35 EGF-like domains. Contains 3 LNR (Lin/Notch) repeats.
Post-translational modificationsSynthesized in the endoplasmic reticulum as an inactive form which is proteolytically cleaved by a furin-like convertase in the trans-Golgi network before it reaches the plasma membrane to yield an active, ligand-accessible form. Cleavage results in a C-terminal fragment N(TM) and a N-terminal fragment N(EC). Following ligand binding, it is cleaved by TNF-alpha converting enzyme (TACE) to yield a membrane-associated intermediate fragment called notch extracellular truncation (NEXT). This fragment is then cleaved by presenilin dependent gamma-secretase to release a notch-derived peptide containing the intracellular domain (NICD) from the membrane. Hydroxylated by HIF1AN. Can be either O-glucosylated or O-xylosylated at Ser-613 by POGLUT1.
Cellular localizationCell membrane and Nucleus. Cytoplasm. Following proteolytical processing NICD is translocated to the nucleus. Retained at the cytoplasm by C8orf4 (PubMed:25985737).