Notch1, Intracellular Domain (Notch Homolog Protein 1, hN1, Translocation-associated Notch Protein TAN-1)

Notch1 is a 300kD type I transmembrane glycoprotein that is one of four human Notch homologues involved in developmental processes. Notch signaling is important for maintaining stem cells and inducing differentiation, especially in the nervous system and lymphoid tissues. Notch can specify binary cell fates like promoting T- over B-cell development from a common precursor. > 50% of human T-lineage acute lymphoblastic leukemia (T-ALL) have activating mutations of Notch1. Human Notch1 is synthesized as a 2556aa precursor that contains an 18aa signal sequence, a 1718aa extracellular domain (ECD) with 36 EGF-like repeats and three Lin-12/Notch repeats (LNR), a 23aa transmembrane (TM) segment and a 785aa cytoplasmic domain containing six ankyrin repeats, a glutamine-rich domain and a PEST sequence. The 11th and 12th EGF-like repeats bind ligands including Jagged and Delta-like families in humans. O-fucosylation by Fringe family members within this region can inhibit the interaction of Notch with Jagged ligands, thereby promoting Delta-like ligand interactions. Notch1 receptor undergoes post- translational furin-type proteolytic cleavage, forming a heterodimer through interaction of a hydrophobic area C-terminal to the LNR on the 1647aa ligand-binding extracellular region with the 891aa transmembrane, cytoplasmic portion. Upon ligand binding, additional sequential proteolysis by TNF-converting enzyme (ADAM17) and the presenilin-dependent gamma-secretase results in the release of the Notch intracellular domain (NICD) which translocates into the nucleus, activating transcription of Notch-responsive genes. \n\nApplications: \nSuitable for use in ELISA, Western Blot and Immunocytochemistry. Other applications not tested.\nRecommended Dilutions:\nWestern Blot: 1ug/ml detects Notch 1 in recombinant human Notch1 Fc Chimera.\nImmunocytochemistry: 8-25ug/ml for immersion fixed Saos-2 human osteosarcoma cells.\nOptimal dilutions to be determined by the researcher.\nStorage and Stability:\nLyophilized powder may be stored at -20 degrees C. Stable for 12 months at -20 degrees C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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