Activity staining of glutathione peroxidase after two-dimensional gel electrophoresis

Chang Won Kho; Sung Goo Park; Do Hee Lee; Sayeon Cho; Goo Taeg Oh; Seongman Kang; Byoung Chul Park

Activity staining of glutathione peroxidase after two-dimensional gel electrophoresis

Chang Won Kho, Sung Goo Park, Do Hee Lee, Sayeon Cho, Goo Taeg Oh, Seongman Kang, Byoung Chul Park

Department of Life Sciences

Research output: Contribution to journal › Article › peer-review

6 Citations (Scopus)

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.

Original language	English
Pages (from-to)	369-373
Number of pages	5
Journal	Molecules and cells
Volume	18
Issue number	3
Publication status	Published - 2004 Dec 31

Keywords

2-D gel
Activity staining
Glutathione
Glutathione peroxidase

ASJC Scopus subject areas

Molecular Biology
Cell Biology

Cite this

@article{d8149db8534947d49e5074acb5e432b0,

title = "Activity staining of glutathione peroxidase after two-dimensional gel electrophoresis",

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.",

keywords = "2-D gel, Activity staining, Glutathione, Glutathione peroxidase",

author = "Kho, {Chang Won} and Park, {Sung Goo} and Lee, {Do Hee} and Sayeon Cho and Oh, {Goo Taeg} and Seongman Kang and Park, {Byoung Chul}",

year = "2004",

month = dec,

day = "31",

language = "English",

volume = "18",

pages = "369--373",

journal = "Molecules and Cells",

issn = "1016-8478",

publisher = "Korean Society for Molecular and Cellular Biology",

number = "3",

}

TY - JOUR

T1 - Activity staining of glutathione peroxidase after two-dimensional gel electrophoresis

AU - Kho, Chang Won

AU - Park, Sung Goo

AU - Lee, Do Hee

AU - Cho, Sayeon

AU - Oh, Goo Taeg

AU - Kang, Seongman

AU - Park, Byoung Chul

PY - 2004/12/31

Y1 - 2004/12/31

N2 - 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.

AB - 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.

KW - 2-D gel

KW - Activity staining

KW - Glutathione

KW - Glutathione peroxidase

UR - http://www.scopus.com/inward/record.url?scp=14544304561&partnerID=8YFLogxK

M3 - Article

C2 - 15650335

AN - SCOPUS:14544304561

SN - 1016-8478

VL - 18

SP - 369

EP - 373

JO - Molecules and Cells

JF - Molecules and Cells

IS - 3

ER -

Activity staining of glutathione peroxidase after two-dimensional gel electrophoresis

Abstract

Keywords

ASJC Scopus subject areas

Other files and links

Fingerprint

Cite this