Solubilization of proteins in 2DE: an outline.
Protein solubilization for two-dimensional electrophoresis (2DE) has to break molecular interactions to separate the biological contents of the material of interest into isolated and intact polypeptides. This must be carried out in conditions compatible with the first dimension of 2DE, namely isoelectric focusing. In addition, the extraction process must enable easy removal of any nonprotein component interfering with the isoelectric focusing. The constraints brought in this process by the peculiar features of isoelectric focusing are discussed, as well as their consequences in terms of possible solutions and limits for the solubilization process.
Rabilloud
Thierry
T
CEA-DSV-iRTSV/LBBSI and UMR CNRS 5092, CEA Grenoble, 17 rue des martyrs, F-38054 Grenoble Cedex 9, France.
D004220:Disulfides / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D007525:Isoelectric Focusing / Q000379:methods*; D007526:Isoelectric Point; D011506:Proteins / Q000737:chemistry*; D012995:Solubility
D004220:Disulfides; D011506:Proteins