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

Autor:	Bjørn J. Bolann, R J Ulvik
Rok vydání:	1991
Předmět:	chemistry.chemical_classification Chromatography Antioxidant biology medicine.medical_treatment Biochemistry (medical) Clinical Biochemistry Superoxide dismutase activity Superoxide dismutase Enzyme Biochemistry chemistry medicine biology.protein Dismutase Quantitative analysis (chemistry)
Zdroj:	Clinical Chemistry. 37:1993-1999
ISSN:	1530-8561 0009-9147
DOI:	10.1093/clinchem/37.11.1993
Popis:	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.
Databáze:	OpenAIRE
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