title: Isoelectric focusing of high-molecular-weight protein complex under native conditions using agarose gel.
abstract: The isolation and characterization of protein complexes are essential steps toward understanding cellular functions. A method for separating and characterizing high-molecular-weight protein complexes using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) with native agarose gel isoelectric focusing (IEF) is described. Using this method, fractions containing high-molecular-weight protein complexes were analyzed. The advantages of using native agarose gel IEF include the ability to concentrate the protein complexes and the ease of handling when performing 2D separations. Although limited with respect to the size of molecules and particles that may be separated, this method is useful for the isolation and characterization of high-molecular-weight protein complexes.
authors.affiliation: Supramolecular Biology, International Graduate School of Arts and Sciences, Yokohama City University, Suehiro-cho 1-7-29, Tsurumi-ku, Yokohama 230-0045, Japan.
authors.forename: Ryo
authors.initials: R
authors.lastname: Yokoyama
authors.forename: Yuko
authors.initials: Y
authors.lastname: Iwafune
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kawasaki
authors.forename: Hisashi
authors.initials: H
authors.lastname: Hirano
mesh_terms: D004587:Electrophoresis, Agar Gel / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D007525:Isoelectric Focusing / Q000379:methods*; D008970:Molecular Weight; D046912:Multiprotein Complexes / Q000302:isolation & purification*; D046988:Proteasome Endopeptidase Complex / Q000302:isolation & purification; D040901:Proteomics / Q000379:methods; D029701:Saccharomyces cerevisiae Proteins / Q000302:isolation & purification
chemical_list: D046912:Multiprotein Complexes; D029701:Saccharomyces cerevisiae Proteins; D046988:Proteasome Endopeptidase Complex