A simplified procedure for purification of cytochrome P-450 by preparative ampholine gel for isoelectric focusing.
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.
Kuwada
M
M
Department of Biochemistry, School of Medicine, Kitasato University, Sagamihara, Kanagawa, Japan.
Maki
J
J
Hasumi
H
H
Furudate
S
S
Takahasi
K
K
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
D000596:Amino Acids; D000665:Ampholyte Mixtures; D011073:Polyamines; D011108:Polymers; C002023:Ampholine; D003577:Cytochrome P-450 Enzyme System