Evaluation of negative staining technique for determination of CN--insensitive superoxide dismutase activity

Guglielmo De Rosa, David S. Duncan, Carl L Keen, Lucille S. Hurley

Research output: Contribution to journalArticle

20 Scopus citations

Abstract

The different forms of superoxide dismutase (superoxide-superoxide oxido-reductase, EC 1.15.1.1) have been studied, in tissues of rat, mouse and chicken, by the electrophoresis-nitro blue tetrazolium technique proposed by Beauchamp. Similar enzyme patterns were evident in every tissue. A fast migrating CN--sensitive form of dismutase activity was present in isolated liver mitochondria of each species. Chicken and mouse liver mitochondria, as well as whole homogenate of every tissue of these two species, showed two additional slow-migrating bands of CN--insensitive activity. In contrast, such bands were not detectable in mitochondria isolated from rat liver or in any of the rat tissues analyzed by this technique. Prior to their electrophoretic separation, the samples were analyzed for CN--insensitive superoxide dismutase activity by a spectrophotometric assay; by this assay it was possible to demonstrate and quantitate a CN--insensitive superoxide dismutase activity in every preparation. Two units of CN--insensitive activity were applied to the gels for each sample. These results indicate that the electrophoresis-nitro blue tetrazolium technique is unsuitable for the detection of the rat CN--insensitive form of superoxide dismutase in crude preparations such as whole tissue homogenates or isolated mitochondria.

Original languageEnglish (US)
Pages (from-to)32-39
Number of pages8
JournalBBA - Enzymology
Volume566
Issue number1
DOIs
StatePublished - Jan 12 1979

Keywords

  • (Isoenzyme)
  • CN-insensitive
  • Negative staining
  • Superoxide dismutase

ASJC Scopus subject areas

  • Medicine(all)

Fingerprint Dive into the research topics of 'Evaluation of negative staining technique for determination of CN<sup>-</sup>-insensitive superoxide dismutase activity'. Together they form a unique fingerprint.

  • Cite this