Comparison of solubilization and preparation procedures for membrane-enriched proteome analysis

Abstract

Proteome analysis of membrane proteins by two-dimentional electrophoresis (2-DE) is still insufficient due to the problem of membrane proteins solubilization. Additionally, nucleic acids and lipids are known to interfere with the profile on 2-DE, and removal of these substances is often required. In this study, three reagents from Bio-X Inc. were used to remove these interfering substances from protein. Prior to loading, samples were treated with the reagents for 30-60 min at 30°C. Pretreatment with the reagents led to high-resolution separations of proteins. Here, we describe how sample pretreatment using the reagents improved proteomic maps of proteins extracted from Escherichia coli and Saccharomyces cerevisiae. To remove lipids enhanced the solubility and separation of membrane proteins from cells.