A highly sensitive method for determining both Mn- and CuZn superoxide dismutase activities in tissues and blood cells

Minoru Nakano, Hirokazu Kimura, Masanori Hara, Minoru Kuroiwa, Masahiko Kato, Kazuhito Totsune, Toshikazu Yoshikawa

Research output: Contribution to journalArticlepeer-review

58 Citations (Scopus)

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 O2- 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.

Original languageEnglish
Pages (from-to)277-280
Number of pages4
JournalAnalytical Biochemistry
Volume187
Issue number2
DOIs
Publication statusPublished - 1990 Jun

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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