Microtiter plate assay for superoxide dismutase using MTT reduction by superoxide

Madesh Muniswamy, K. A. Balasubramanian

Research output: Contribution to journalArticle

282 Citations (Scopus)

Abstract

A simple microtiter plate based colorimetric assay for superoxide dismutase is described. The method, involves generation of superoxide by pyrogallol autoxidation and the inhibition of superoxide dependent reduction of the tetrazolium dye MTT [3-(4,5-dimethyl thiazol-2-yl) 2,5-diphenyl tetrazolium bromide] to its formazan, measured at 570 nm. The reaction was terminated by the addition of dimethyl sulfoxide (DMSO) which also helps to solubilize the formazan formed and the colour evolved was stable for many hours. The method was compared with other known methods to measure the activity of purified erythrocyte Cu,ZnSOD and superoxide dismutase activity from various rat tissues. This procedure involves inexpensive reagents, allows a rapid and sensitive measurement of SOD activity and the microtiter plate assay is suitable for use with large number of samples.

Original languageEnglish (US)
Pages (from-to)184-188
Number of pages5
JournalIndian Journal of Biochemistry and Biophysics
Volume35
Issue number3
StatePublished - Jun 1 1998
Externally publishedYes

Fingerprint

Formazans
Superoxides
Superoxide Dismutase
Assays
Pyrogallol
Dimethyl Sulfoxide
Bromides
Rats
Coloring Agents
Tissue
Color
Erythrocytes
diphenyl

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics

Cite this

Microtiter plate assay for superoxide dismutase using MTT reduction by superoxide. / Muniswamy, Madesh; Balasubramanian, K. A.

In: Indian Journal of Biochemistry and Biophysics, Vol. 35, No. 3, 01.06.1998, p. 184-188.

Research output: Contribution to journalArticle

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