Abstract
To enable two-dimensional (2-D) gel electrophoresis with high reproducibility, we have recently developed and reported a new procedure by which a whole lysate from Arabidopsis can be directly subjected to isoelectric focusing (IEF) without any desalting steps. In this study, we applied the IEF method to the 2-D gel electrophoresis of membrane proteins without removing SDS from samples. Plasma membrane enriched fractions were isolated using a two-phase partition system. Fractions were then dissolved by a solution with or without 1.5% SDS, and then separated by the 2-D gel electrophoresis using the IEF method. Each of protein samples gave a high-resolution pattern on the 2-D gel. In samples dissolved with solution containing 1.5% SDS, some proteins were extracted with higher efficiency. Therefore, we show that more membrane proteins were quantitatively detectable on 2-D gel without any desalting steps by using the IEF method.
