S-nitrosoproteome in endothelial cells revealed by a modified biotin swap method coupled with Western blot-based two-dimensional gel electrophoresis.
Native electrophoresis and western blot evaluation: methodology and functions.
Characterization of human insulin-like progress factor-binding proteins by two-dimensional polyacrylamide gel electrophoresis and Western ligand blot evaluation.
The insulin-like progress issue (IGF)-binding proteins (IGFBPs) from grownup human serum, amniotic fluid, and cerebrospinal fluid have been analyzed by a modified two-dimensional gel electrophoresis adopted by Western ligand blotting. The samples have been subjected to immobilized pH gradient isoelectric focusing within the first dimension, adopted by nondenaturing SDS-PAGE within the second dimension and autoradiography after ligand blotting with [125I]IGF-I or [125I]IGF-II.
The identification of the binding proteins was confirmed by immunoblotting and immunoprecipitation with particular antibodies. Utilizing this methodology, all six human excessive affinity IGFBPs could possibly be clearly separated from one another in keeping with their molecular mass and isoelectric factors (pI). All IGFBPs exhibited a wide range of particular pI isoforms, which presumably signify posttranslational modifications.