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Measurement of superoxide dismutase, catalase and glutathione peroxidase in cultured cells and tissue.
Nat Protoc. 2010 Jan; 5(1):51-66.NP

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.

Authors+Show Affiliations

Department of Molecular Physiology and Biophysics, The University of Iowa Carver College of Medicine, Iowa City, Iowa, USA.No affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

20057381

Citation

Weydert, Christine J., and Joseph J. Cullen. "Measurement of Superoxide Dismutase, Catalase and Glutathione Peroxidase in Cultured Cells and Tissue." Nature Protocols, vol. 5, no. 1, 2010, pp. 51-66.
Weydert CJ, Cullen JJ. Measurement of superoxide dismutase, catalase and glutathione peroxidase in cultured cells and tissue. Nat Protoc. 2010;5(1):51-66.
Weydert, C. J., & Cullen, J. J. (2010). Measurement of superoxide dismutase, catalase and glutathione peroxidase in cultured cells and tissue. Nature Protocols, 5(1), 51-66. https://doi.org/10.1038/nprot.2009.197
Weydert CJ, Cullen JJ. Measurement of Superoxide Dismutase, Catalase and Glutathione Peroxidase in Cultured Cells and Tissue. Nat Protoc. 2010;5(1):51-66. PubMed PMID: 20057381.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Measurement of superoxide dismutase, catalase and glutathione peroxidase in cultured cells and tissue. AU - Weydert,Christine J, AU - Cullen,Joseph J, Y1 - 2009/12/17/ PY - 2010/1/9/entrez PY - 2010/1/9/pubmed PY - 2010/3/23/medline SP - 51 EP - 66 JF - Nature protocols JO - Nat Protoc VL - 5 IS - 1 N2 - 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. SN - 1750-2799 UR - https://www.unboundmedicine.com/medline/citation/20057381/Measurement_of_superoxide_dismutase_catalase_and_glutathione_peroxidase_in_cultured_cells_and_tissue_ L2 - https://doi.org/10.1038/nprot.2009.197 DB - PRIME DP - Unbound Medicine ER -