Application of sequential extraction procedures and glycoprotein blotting for the characterization of the 2-D polypeptide patterns of barley seed proteins.
Barley (Hordeum vulgare L.) proteins were sequentially extracted from ground seeds with Tris-HCl buffer, 55% 2-propanol, 55% 2-propanol containing 1% dithiothreitol, and 6 M urea containing 2% Nonidet P-40 and 1% dithiothreitol. The protein composition of these solubility fractions was then analyzed by high resolution two-dimensional gel electrophoresis with immobilized pH gradient 4-9 in the first dimension, followed by silver staining and glycoprotein blotting, respectively, for a more detailed characterization of the two-dimensional polypeptide pattern of barley seed proteins.
D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006023:Glycoproteins / Q000302:isolation & purification; D001467:Hordeum / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D010940:Plant Proteins / Q000302:isolation & purification*; D012639:Seeds / Q000737:chemistry; D012995:Solubility; D013194:Staining and Labeling
D006023:Glycoproteins; D010940:Plant Proteins