Notch4 (Notch homolog 4, Drosophila, INT3)

Notch4 is a 250kD member of the NOTCH family of proteins. It is primarily expressed on endothelium. Human Notch4 is a 1980aa type I transmembrane (TM) glycoprotein that contains a 1424 aa extracellular domain (ECD) and a 535aa cytoplasmic region. Notch4 undergoes proteolytic processing at multiple sites. There is probable cleavage of the ECD by furin in the Golgi between aa1310-1410. This creates a mature, potentially disulfide-linked heterodimer at the cell surface. Upon ligand binding, two sequential cleavages occur, one in the ECD after Ala1431, and a second in the TM domain after Lys1466. This creates a soluble cytoplasmic fragment that interacts with RBP-Jkappa and activates genes associated with HES-1. Over cytoplasmic aa1778-2003, human Notch-4 is 78% aa identical tomouse Notch4. Applications: Suitable for use in Immunocytochemistry, ELISA and Western Blot. Other applications not tested. Recommended Dilution: Immunocytochemistry: 10ug/ml, ELISA (Direct): 0.5-1ug/ml. Detection limit ~2ng/well., Western Blot: 1-2ug/ml. Detection limit ~5ng/lane under non-reducing and reducing conditions., Optimal dilutions to be determined by the researcher. Storage and Stability: Lyophilized powder may be stored at 4°C for short-term only. Reconstitute to nominal volume by adding sterile 40-50% glycerol, PBS and store at -20°C. Reconstituted product is stable for 12 months at -20°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.