Abstract
A highly sensitive method for determining the superoxide dismutase (EC 1.15.1.1) in various tissues and blood cells is described. This method involves inhibition of a cypridina luciferin analog that is chemiluminescence dependent upon O 2 − generated by hypoxanthinexanthine oxidase. Manganeous superoxide dismutase, which is sensitive to sodium dodecyl sulfate, was determined and calculated by subtraction of superoxide dismutase activity in tissue extract treated with this detergent (CuZn superoxide dismutase) from that in untreated tissue extract (total superoxide dismutase). Both Mn- and CuZn superoxide dismutase activities were expressed as equivalent nanograms of bovine erythrocyte superoxide dismutase per milliliter. Sensitivity limits of the chemiluminescence methods with 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo[1,2- a]pyrazin-3-one and 2-methyl-6-phenyl-3,7-dihydroimidazo[1,2- a]pyrazin-3-one as cypridina luciferin analogs were 1 ng and 2–3 ng of superoxide dismutase/ml, respectively.
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