High Speed Isoelectric Focusing of Proteins Enabling Rapid Two-Dimensional Gel Electrophoresis

While two-dimensional electrophoresis (2-DE) of proteins, as we know it, was first described nearly four decades ago (O’Farrell, 1975), it continues to be a critical component of comprehensive proteomics analysis today. Continuing refinements of 2-DE, particularly the development of immobilized pH gradients (Righetti, 1990), new chaotropes (Vecchio et al. 1984, Rabilloud 1998) and detergents (Seddon 2004, Rabilloud et al. 1999) that increase and maintain the solubility of hydrophobic proteins, and sample fractionation strategies (Smejkal and Lazarev 2005, Di Girolamo et al. 2011, Boschetti et al. 2007) that lessen the complexity of proteomes, have further increased the utility of the method.