Improvement of a direct spectrophotometric assay for routine determination of superoxide dismutase activity.

[摘要] The growing interest in measuring superoxide dismutase (EC 1.15.1.1) in many diseases calls for useful routine assays. For this purpose, the direct spectrophotometric method of Marklund (J Biol Chem 1976;251:7504-7) was improved to offer an alternative to the imprecise, indirect assays currently used. The decay of O2.- (from KO2) at pH 9.5 was monitored as the decrease in delta A (delta A = A250nm-A360nm). Superoxide dismutase was determined from the pseudo-first-order rate constant of O2.- dismutation. The precision of the assay was improved by increasing the concentration of O2.- and expanding the interval for measurements of O2.- concentrations to 4-16 mumol/L. Other assay characteristics, including temperature, were also optimized. In hemolysate the assay had a within-day CV of 5.5-13% and a between-day CV of 4%. Mn-superoxide dismutase and some superoxide dismutase mimics are inhibited at alkaline pH. Therefore, the method is primarily recommended for Cu,Zn-superoxide dismutase.

[学科分类] 过敏症与临床免疫学