title: Two-dimensional gel electrophoresis: glass tube-based IEF followed by SDS-PAGE.
abstract: The genome information combined with data derived from modern mass spectrometry enables us to determine the identity of a protein once it is isolated from a complex mixture. Two-dimensional gel electrophoresis established more than three decades ago serves as a powerful protocol to isolate many proteins at once for such protein analysis. In the first two decades, the original procedure to use a glass tube-based isoelectric focusing (IEF) had been commonly used. Since an IEF in glass tubes is rather difficult to maneuver, a new method to use an IEF on a thin agarose slab backed by a plastic film (IPG Dry Strip) has been invented and is now widely used. In this chapter, we describe the original protocol that uses a glass tube-based IEF because, the capacity of protein loading and resolving power of this type of classic two-dimensional gel is still indispensible.
authors.affiliation: University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA. hiroyuki-matsumoto@ouhsc.edu
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Matsumoto
authors.forename: Hisao
authors.initials: H
authors.lastname: Haniu
authors.forename: Biji T
authors.initials: BT
authors.lastname: Kurien
authors.forename: Naoka
authors.initials: N
authors.lastname: Komori
mesh_terms: D000818:Animals; D002021:Buffers; D004587:Electrophoresis, Agar Gel / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006801:Humans; D007525:Isoelectric Focusing / Q000379:methods; D011489:Protein Denaturation; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*; D014508:Urea / Q000737:chemistry
chemical_list: D002021:Buffers; D011506:Proteins; D014508:Urea