Author:

Keywords:

Science & Technology, Life Sciences & Biomedicine, Biochemical Research Methods, Biochemistry & Molecular Biology, Cell Biology, Notch signaling, Notch cleavage, NICD fragment, Cell-based assay, S3 cleavage, gamma-Secretase, Immunoblot, DOMAIN, Amyloid Precursor Protein Secretases, Animals, Blotting, Western, HEK293 Cells, Humans, Mice, Proteolysis, Receptors, Notch, Transfection, 0399 Other Chemical Sciences, 0601 Biochemistry and Cell Biology, Developmental Biology, 3101 Biochemistry and cell biology, 3404 Medicinal and biomolecular chemistry

Abstract:

With the increasing appreciation of the role of Notch in development and disease, measuring its cleavage and signaling activity in cellular systems has become important. Here we describe a cell-based method to analyze the cleavage of Notch at the S3 site by γ-secretase. HEK cells are transfected with an N-terminal truncated and myc-labeled mNotchΔE construct which can be easily and quantitatively detected by western blotting.