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Title 

Activity staining of glutathione peroxidase after two-dimensional gel electrophoresis

Authors 

Chang Won KhoSung Goo ParkDo Hee LeeS ChoG T OhS KangByoung Chul Park

Publisher 

Springer Verlag (Germany)

Issue Date 

2004

Citation 

Molecules and Cells, vol. 18, no. 3, pp. 369-373

Keywords 

2-D gelactivity stainingglutathioneglutathione peroxidaseenzyme activitystainingtwo dimensional gel electrophoresiselectrophoresis, polyacrylamide gelglutathione transferasesilver staining

Abstract 

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). After this proteins with GPx activity appeared as clear zones on a purple background. This relatively simple activity staining method could be useful for rapid screening of proteins with GPx activity in cell extracts.

ISSN 

1016-8478

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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