Journal
ELECTROPHORESIS
Volume 26, Issue 14, Pages 2809-2822Publisher
WILEY
DOI: 10.1002/elps.200500060
Keywords
convective interaction media monoliths; plasma membranes; protein mapping; selective extraction
Funding
- NCI NIH HHS [CA42714] Funding Source: Medline
- NCRR NIH HHS [P20RR017695] Funding Source: Medline
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A model system for selective solubilization and fast separation of proteins from the rat liver membrane fraction and purified rat liver plasma membranes for their further proteomic analysis is presented. For selective solubilization, high-pH solutions and a concentrated urea solution, combined with different detergents, are used. After extraction, proteins are separated by anion-exchange chromatography or a combination of anion- and cation-exchange chromatography with convective interaction monolithic supports. This separation method enables fast and effective prefractionation of membrane proteins based on their hydrophobicity and charge prior to one-dimensional (1-D) and 2-D electrophoresis and mass spectrometry. By use of this sample preparation method, the less-abundant proteins can be detected and identified.
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