title: Two-step OFFGEL approach for effective peptide separation compatible with iTRAQ labeling.
abstract: Shotgun proteomic analyses are increasingly becoming methods of choice for complex samples. The development of effective methods for fractionating peptides to reduce the complexity of the sample before mass analysis is a key point in this strategy. The OFFGEL technology has recently become a tool of choice in proteomic analysis at peptide level. This OFFGEL electrophoresis (OGE) approach allows the in-solution separation of peptides from various biological sources by isoelectric focusing in highly resolved 24 fractions. It was also demonstrated that OGE technology is a filtering tool for pI-based validation of peptide identification. As peptide OGE is compatible with iTRAQ labeling, OGE is finding valuable applications in quantitative proteomics as well. The aim of this study is to explain a new 2D-OGE approach that improves the proteomic coverage of complex mixtures such as colorectal cell line lysates, and which is compatible with iTRAQ labeling.
authors.affiliation: Paul Papin Cancer Center, Institut de Cancérologie de l'Ouest, INSERM U892, Angers, France.
authors.forename: Alexis
authors.initials: A
authors.lastname: Rideau
authors.forename: Damien
authors.initials: D
authors.lastname: Besson
authors.forename: Alice
authors.initials: A
authors.lastname: Boissard
authors.forename: Olivier
authors.initials: O
authors.lastname: Coqueret
authors.forename: Catherine
authors.initials: C
authors.lastname: Guette
mesh_terms: D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D007553:Isotope Labeling / Q000379:methods*; D010455:Peptides / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D020543:Proteome / Q000032:analysis* / Q000737:chemistry; D040901:Proteomics / Q000379:methods; D053719:Tandem Mass Spectrometry
chemical_list: D010455:Peptides; D020543:Proteome
keywords: OFFGEL; OGE; Technology; iTRAQ