title: No peptide left behind: the "out of range" recovery in IPG-IEF fractionation.
abstract: IEF is often used in multidimensional shotgun proteomics and the narrow range of 3.5-4.5 is the recommended pH interval for the fractionation of tryptic peptides. Usually, even if IEF is performed in IPG strip with a narrow range pH, the entire sample must be loaded onto the strip, including the "out of IPG range" peptides. We describe a simple protocol to recover at least a part of these missing peptides and show that this recovery significantly influences the overall fractionation result, increasing the number of the identified proteins and the protein coverage.
authors.affiliation: Department of Medicine, University of Padova, Via Giustiniani 2, 35121, Padua, Italy.
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Iori
authors.forename: Marcello
authors.initials: M
authors.lastname: Rattazzi
authors.forename: Renato
authors.initials: R
authors.lastname: Millioni
mesh_terms: D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing / Q000295:instrumentation / Q000379:methods*; D010455:Peptides / Q000737:chemistry*; D040901:Proteomics / Q000295:instrumentation / Q000379:methods
chemical_list: D010455:Peptides