title: Redox options in two-dimensional electrophoresis.
abstract: Two-dimensional electrophoresis is usually run on fully reduced samples. Under these conditions even covalently bound oligomers are dissociated and individual polypeptide chains may be fully unfolded by both, urea and SDS, which maximizes the number of resolved components and allows their pI and M(r) to be most accurately evaluated. However, various electrophoretic protocols for protein structure investigation require a combination of steps under varying redox conditions. We review here some of the applications of these procedures. We also present some original data about a few related samples -- serum from four species: Homo sapiens, Mus musculus, Rattus norvegicus, Bos taurus -- which we run under fully unreduced and fully reduced conditions as well as with reduction between first and second dimension. We demonstrate that in many cases the unreduced proteins migrate with a better resolution than reduced proteins, mostly in the crowded 'alpha-globulin' area of pI 4.5-6 and M(r) 50-70 kDa.
authors.affiliation: Kennedy Institute of Rheumatology Division, Faculty of Medicine, Imperial College London, United Kingdom.
authors.forename: R
authors.initials: R
authors.lastname: Wait
authors.forename: S
authors.initials: S
authors.lastname: Begum
authors.forename: D
authors.initials: D
authors.lastname: Brambilla
authors.forename: A M
authors.initials: AM
authors.lastname: Carabelli
authors.forename: F
authors.initials: F
authors.lastname: Conserva
authors.forename: A
authors.initials: A
authors.lastname: Rocco Guerini
authors.forename: I
authors.initials: I
authors.lastname: Eberini
authors.forename: R
authors.initials: R
authors.lastname: Ballerio
authors.forename: M
authors.initials: M
authors.lastname: Gemeiner
authors.forename: I
authors.initials: I
authors.lastname: Miller
authors.forename: E
authors.initials: E
authors.lastname: Gianazza
mesh_terms: D000818:Animals; D002417:Cattle; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D007525:Isoelectric Focusing / Q000379:methods*; D051379:Mice; D010084:Oxidation-Reduction; D011506:Proteins / Q000032:analysis*; D051381:Rats
chemical_list: D011506:Proteins