Determination of Manganese Superoxide Dismutase Activity By Direct Spectrophotometry |
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| Authors: | Bj rn J. Bolann Arild Tanger s Rune J. Ulvik |
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| Affiliation: | a Section of Biochemistry, Institute of Clinical Biology, University of Bergen, Bergen, Norwayb Institute of Biochemistry and Molecular Biology, University of Bergen, Bergen, Norway |
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| Abstract: | A method to determine Mn-superoxide dismutase activity by measuring directly the rate of decay of O2- in a spectrophotometer, is described. Decay of O2- generated by KO2 at pH 9.5, was monitored as the fall in absorbance (A250nm-A360nm). Mn-superoxide dismutase was determined as the activity of cyanide-resistant superoxide dismutase, calculated from the rate of O2- dismutation. Mn-superoxide dismutase could be determined in the presence of a 700 times higher Cu, Zn-superoxide dismutase activity. The alkaline pH did not cause analytical problems. The assay was used to measure both Mn- and Cu, Zn-superoxide dismutase activity in mitochondrial preparations. The assay had a detection limit of 2.8 ng/ml when Mn-superoxide dismutase from E. coli was used, and the between-day CV was 5.8%. The assay is an alternative to indirect methods for detecting superoxide dismutase activity. |
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| Keywords: | Superoxide dismutase Manganese Spectrophotometry |
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