Activity Staining of Glutathione Peroxidase after Two-dimensional Gel Electrophoresis
Kho, C.W.;Park, S.G.;Lee, D.H.;Park, B.C.(Korea Research Institute of Bioscience and Biotechnology, Daejeon, Republic of Korea)E-mail:parkbc@kribb.re.kr | Cho, S.Y.(Chung-Ang University, Seoul, Republic of Korea) | Oh, G.T.(Ewha Womans University, Seoul, Republic of Korea) | Kang, S.M.(Korea University, Seoul, Republic of Korea)
We have developed a method for rapid activity staining of proteins with glutathione peroxidase (GPx) activity after 2-D gel electrophoresis. After separating proteins extracted from yeast, or mouse red blood cells, by two-dimensional gel electrophoresis, SDS was removed and the gel was submerged in a Tris-HCl buffer containing glutathione and hydrogen peroxide, followed by incubation with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and phenazine methosulfate (PMS).
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