The optimized conditions of two-dimensional polyacrylamide gel electrophoresis for proteomic analysis of Saccharomyces cerevisiae

Proteome analysis is most commonly based on a two-dimensional gel electrophoresis to separate and visualize proteins for protein identification. Although this technique is powerful, sensitive, and mature, doubts remain concerning its ability to characterize all the elements of a proteome. For 2-D gel analysis, it is very important to obtain highest possible resolution in protein separation, especially for crowded areas on gel. In this study, we describe an approach for the accurate quantification of protein spots reducing background noise on gel. The proteome of S. cerevisiae was analysed by 2-D gel electrophoresis, varying existing conventional procedures to obtain maximum high quality protein spots on gel. The modifications in method clearly improve the resolution of protein identification spots as compared to conventional methods.