C-NAAERTPSHSGHLQaa 2396-2409 (the total protein is 2471 aa).
Synthesized 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 Functions 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 RBP-J kappa and activates genes of the enhancer of split locus. Affects the implementation of differentiation, proliferation and apoptotic programs.
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|WB||1/400. Detects a band of approximately 110 kDa (predicted molecular weight: 110 kDa).|
Western Blot analysis for Notch 2 expression performed on 100
Lanes1: 24 hours after transfection.
Lane 2: 48 hours after transfection.
Lane 3: 72 hours after transfection.
Lane 4: Control cells 24 hours after transfection.
Lane 5: Control cells 48 hours after transfection.
Lane 6: Control cells 72 hours after transfection.
The band at about 110kD represents active Notch 2. This band is not seen in the control cells, confirming that it is Notch 2. The intracellular domain of Notch 2 has a predicted band size of 110kD, corresponding to this band.
Protein cell lysates were run on a 10% SDS-page gel, blotted onto Hybond C membrane, blocked overnight in PBS-Tween 20 supplemented with 5% Non-fat Milk and hybridized with ab8927 rabbit polyclonal antibody specific for Notch 2 at a 1:400 dilution.
ECL was used as visualization method.