Superoxide dismutase micro assay in biological material.

Hoppe-Seyler's Zeitschrift für physiologische Chemie
W Bohnenkamp, U Weser

Abstract

A micro assay for the rapid and convenient determination of superoxide dismutase activity in limited amounts of biological material was devised and successfully employed. The combination of the formazan derivative colour formation induced by reaction of O2theta with nitroblue tetrazolium and a suitable analytical polyacrylamide gel electrophoresis system was used. It was possible to show that the reactivity of soluble superoxide dismutases in polyacrylamide gels was proportional to the enzyme concentrations employed. Bovine erythrocyte Cu, Zn-superoxide dismutase (EC 1.15.1.1) (erythrocuprein) served as a standard throughout. To measure the degree of superoxide dismutase activity, a gel-scanning apparatus was usedThe integrated scanning curve of the unstained portions of the gel proved linearly proportional to the logarithm of the superoxide dismutase activity in the range between 10(-12) and 7 X 10(-11) mol of the bovine enzyme. Although the absolute integral is dependent on the different staining conditions, the slope of the superoxide dismutase calibration curve is highly reproducible. Superoxide dismutase added to crude liver and brain homogenates could be fully detected using this assay. Thus, biological material including...Continue Reading

Citations

Jan 1, 1979·Journal of Neuroscience Research·R Fried
Jun 1, 1977·Biulleten' eksperimental'noĭ biologii i meditsiny·N N Vol'skiĭ, V M Mishin
Feb 1, 1978·Experimental and Molecular Pathology·V M MishinV V Lyakhovich
Oct 1, 1978·Photochemistry and Photobiology·W BorsC Frenzel
Sep 20, 1996·Journal of Chromatography. B, Biomedical Applications·W P Michalski
Sep 24, 1976·Biochimica Et Biophysica Acta·W Bohnenkamp, U Weser

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