Electrophoresis · 1998 · 330 citations · 24 references
Life SciencesBiochemistryMolecular BiologyMembrane BiochemistryProtein BiochemistryProteomicsMembrane ProteinsMembrane ProteomicsProtein ElectrophoresisProtein PurificationProtein SeparationDetergentsHigh-resolution ElectrophoresisMembrane Protein SeparationMembrane Protein SolubilizationSodium Dodecyl Sulfate-polyacrylamide Gel ElectrophoresisTwo-dimensional ElectrophoresisTwo-dimensional Gel Electrophoresis
Severe quantitative loss of protein is often observed in high-resolution two-dimensional electrophoresis of membrane proteins, while the resolution is usually not affected. To improve the solubility of proteins in this technique, we tested denaturing cocktails containing various detergents and chaotropes. Best results were obtained with a denaturing solution containing urea, thiourea, and zwitterionic detergents, synthesized for this purpose. Among the dozen detergents synthesized and tested, amidosulfobetaines with an alkyl tail containing 14-16 carbons proved most efficient, solubilizing previously undetected membrane proteins.
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Mass Spectrometric Sequencing of Proteins from Silver-Stained Polyacrylamide Gels
Andrej Shevchenko, Matthias Wilm, Ole Vorm et al. · Analytical Chemistry · 1996 · 9.1K citations
Thierry Rabilloud, Cecyl Valette, Jean Jacques Lawrence · Electrophoresis · 1994 · 365 citations