Measurement of superoxide dismutase, catalase and glutathione peroxidase in cultured cells and tissue

Christine J Weydert; Joseph J Cullen

doi:10.1038/nprot.2009.197

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Measurement of superoxide dismutase, catalase and glutathione peroxidase in cultured cells and tissue

Journal article

Peer reviewed

Measurement of superoxide dismutase, catalase and glutathione peroxidase in cultured cells and tissue

Christine J Weydert and Joseph J Cullen

Nature protocols, Vol.5(1), pp.51-66

01/2010

DOI: 10.1038/nprot.2009.197

PMCID: PMC2830880

PMID: 20057381

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Abstract

Cells contain a large number of antioxidants to prevent or repair the damage caused by reactive oxygen species, as well as to regulate redox-sensitive signaling pathways. General protocols are described to measure the antioxidant enzyme activity of superoxide dismutase (SOD), catalase and glutathione peroxidase. The SODs convert superoxide radical into hydrogen peroxide and molecular oxygen, whereas the catalase and peroxidases convert hydrogen peroxide into water. In this way, two toxic species, superoxide radical and hydrogen peroxide, are converted to the harmless product water. Western blots, activity gels and activity assays are various methods used to determine protein and activity in both cells and tissue depending on the amount of protein required for each assay. Other techniques including immunohistochemistry and immunogold can further evaluate the levels of the various antioxidant enzymes in tissues and cells. In general, these assays require 24-48 h to complete.

Cell Line

Glutathione Peroxidase - metabolism

Tissue Culture Techniques

Hydrogen Peroxide - metabolism

Catalase - metabolism

Immunohistochemistry - methods

Superoxide Dismutase - chemistry

Animals

Glutathione Peroxidase - chemistry

Superoxides - metabolism

Mice

Cell Culture Techniques

Superoxide Dismutase - metabolism

Voltage-Dependent Anion Channels

Catalase - chemistry

Details

Title: Subtitle: Measurement of superoxide dismutase, catalase and glutathione peroxidase in cultured cells and tissue
Creators: Christine J Weydert - Department of Molecular Physiology and Biophysics, The University of Iowa Carver College of Medicine, Iowa City, Iowa, USA
Joseph J Cullen
Resource Type: Journal article
Publication Details: Nature protocols, Vol.5(1), pp.51-66
DOI: 10.1038/nprot.2009.197
PMID: 20057381
PMCID: PMC2830880
NLM abbreviation: Nat Protoc
ISSN: 1754-2189
eISSN: 1750-2799
Publisher: England
Grant note: R21 CA137230 / NCI NIH HHS I01 BX001318 / BLRD VA R21 CA137230-01A1 / NCI NIH HHS CA137230 / NCI NIH HHS
Language: English
Date published: 01/2010
Academic Unit: Surgery; Radiation Oncology
Record Identifier: 9984051980802771

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