High-Resolution two-dimensional electrophoresis of wheat proteins

A high-resolution two-dimensional electrophoretic for separating wheat endosperm proteins is described. Non-equilibrium pH gradient electrophoresis (NEPHGE) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS- PAGE) were used to examine proteins in non-fractioned extracts of wheat flours. NEPHGE was conducted in 100-μl capillary tubes at 3000 V, for a total of 4800 V h. The second dimension, SDS-PAGE, was performed as a slab mini-gel (70 × 80 × 1.0 mm) with constant application of 200 V. Image analysis of the two-dimensional separations of three wheat varieties showed approximately 35 and 41% homology of Centurk 78 and Rodeo varieties, respectively, when compared to a reference (Scout 66).