Superoxide dismutase: Improved assays and an assay applicable to acrylamide gels

Abstract

Nitro blue tetrazolium has been used to intercept O2- generated enzymically or photochemically. The reduction of NBT by O2- has been utilized as the basis of assays for superoxide dismutase, which exposes its presence by inhibiting the reduction of NBT. Superoxide dismutase could thus be assayed either in crude extracts or in purified protein fractions. The assays described are sensitive to ng/ml levels of super-oxide dismutase and were applicable in free solution or on polyacrylamide gels. The staining procedure for localizing superoxide dismutase on polyacrylamide electrophoretograms has been applied to extracts obtained from a variety of sources. E. coli has been found to contain two superoxide dismutases whereas bovine heart, brain, lung, and erthrocytes contain only one. © 1971.