title: Mass estimation of native proteins by blue native electrophoresis: principles and practical hints.
abstract: Blue native electrophoresis is one of the most popular techniques for mass estimation of native membrane proteins, but the use of non-optimal mass markers and acrylamide gels can compromise accuracy and reliability of the results. We present short protocols taking 10-30 min to prepare optimal sets of membrane protein markers from chicken, rat, mouse, and bovine heart. Especially heart materials from local supermarkets or butcher's shops, e.g. chicken or bovine heart, are ideal sources of high mass membrane protein standards. Considerable discrepancies between the migration behavior of membrane and soluble markers suggest using membrane protein markers for mass estimation of membrane proteins. Soluble standard proteins can be used, with some limitations, when soluble proteins are the focus. Principles and general rules for the determination of mass and oligomeric state of native membrane and soluble proteins are elaborated, and potential pitfalls are discussed.
authors.affiliation: Molecular Bioenergetics, Medical School, Goethe-Universität Frankfurt, Theodor-Stern-Kai 7, Haus 26, D-60590 Frankfurt am Main, Germany.
authors.forename: Ilka
authors.initials: I
authors.lastname: Wittig
authors.forename: Tobias
authors.initials: T
authors.lastname: Beckhaus
authors.forename: Zibiernisha
authors.initials: Z
authors.lastname: Wumaier
authors.forename: Michael
authors.initials: M
authors.lastname: Karas
authors.forename: Hermann
authors.initials: H
authors.lastname: Schägger
mesh_terms: D000818:Animals; D002417:Cattle; D002645:Chickens; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D008565:Membrane Proteins / Q000737:chemistry*; D051379:Mice; D008970:Molecular Weight; D009206:Myocardium / Q000737:chemistry; D051381:Rats
chemical_list: D008565:Membrane Proteins