title: A simplified procedure for purification of cytochrome P-450 by preparative ampholine gel for isoelectric focusing.
abstract: The purification process for cytochrome P450 is very complicated, involving five or more column chromatography steps for the final preparation. This paper describes a reduction in the number of the steps; it can be easily purified from pig testis microsomes with improved the yield. As the first step, DEAE-Toyopearl column chromatography is performed only once and then, as the second step, the partially purified cytochrome P450 is completely purified by a preparative Ampholine PAG-plate Gel for Isoelectric Focusing. The combination reduced the purification to a two-step procedure.
authors.affiliation: Department of Biochemistry, School of Medicine, Kitasato University, Sagamihara, Kanagawa, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Kuwada
authors.forename: J
authors.initials: J
authors.lastname: Maki
authors.forename: H
authors.initials: H
authors.lastname: Hasumi
authors.forename: S
authors.initials: S
authors.lastname: Furudate
authors.forename: K
authors.initials: K
authors.lastname: Takahasi
mesh_terms: D000596:Amino Acids; D000665:Ampholyte Mixtures; D000818:Animals; D005591:Chemical Fractionation; D002845:Chromatography; D003577:Cytochrome P-450 Enzyme System / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D007526:Isoelectric Point; D008297:Male; D008861:Microsomes / Q000201:enzymology; D011073:Polyamines; D011108:Polymers; D012995:Solubility; D013552:Swine; D013737:Testis / Q000201:enzymology
chemical_list: D000596:Amino Acids; D000665:Ampholyte Mixtures; D011073:Polyamines; D011108:Polymers; C002023:Ampholine; D003577:Cytochrome P-450 Enzyme System