title: Activity staining of glutathione peroxidase after electrophoresis on native and sodium dodecyl sulfate polyacrylamide gels.
abstract: Glutathione peroxidase (GSH-Px), from commercial bovine erythrocytes or ammonium sulfate fractionations (30-45%, 45-60%, 60-75% and 75-90% saturations) of ginger rhizome, was detected on polyacrylamide gels after native polyacrylamide gel electrophoresis (PAGE) or sodium dodecyl sulfate (SDS)-PAGE. The gel was submerged in a 50 mM Tris-HCl buffer (pH 7.9) containing 13 mM glutathione and 0.004% hydrogen peroxide with gentle shaking for 10-20 min. The GSH-Px activity was stained with a solution containing 1.2 mM 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and 1.6 mM phenazine methosulfate (PMS) for 10 min. The clear zone of GSH-Px activity on a purple background was found in both native and SDS-PAGE gels. This fast and sensitive method can be used in the process of enzyme purification and characterization of mammalian or plant cells.
authors.affiliation: Department of Pharmacognosy, Graduate Institute of Pharmaceutical Sciences, Taipei Medical University, Taiwan.
authors.forename: Chien-Liang
authors.initials: CL
authors.lastname: Lin
authors.forename: Hsien-Jung
authors.initials: HJ
authors.lastname: Chen
authors.forename: Wen-Chi
authors.initials: WC
authors.lastname: Hou
mesh_terms: D000818:Animals; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004912:Erythrocytes / Q000201:enzymology; D020939:Zingiber officinale / Q000201:enzymology; D005979:Glutathione Peroxidase / Q000302:isolation & purification* / Q000378:metabolism; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods / Q000592:standards*
chemical_list: D005979:Glutathione Peroxidase