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Journal of Biosciences

, Volume 13, Issue 3, pp 305–315 | Cite as

Assay of superoxide dismutase activity in animal tissues

  • Anuradha Nandi
  • I. B. Chatterjee
Article

Abstract

Convenient assays for superoxide dismutase have necessarily been of the indirect type. It was observed that among the different methods used for the assay of superoxide dismutase in rat liver homogenate, namely the xanthine-xanthine oxidase ferricytochromec, xanthine-xanthine oxidase nitroblue tetrazolium, and pyrogallol autoxidation methods, a modified pyrogallol autoxidation method appeared to be simple, rapid and reproducible. The xanthine-xanthine oxidase ferricytochromec method was applicable only to dialysed crude tissue homogenates. The xanthine-xanthine oxidase nitroblue tetrazolium method, either with sodium carbonate solution, pH 10.2, or potassium phosphate buffer, pH 7·8, was not applicable to rat liver homogenate even after extensive dialysis. Using the modified pyrogallol autoxidation method, data have been obtained for superoxide dismutase activity in different tissues of rat. The effect of age, including neonatal and postnatal development on the activity, as well as activity in normal and cancerous human tissues were also studied. The pyrogallol method has also been used for the assay of iron-containing superoxide dismutase inEscherichia coli and for the identification of superoxide dismutase on polyacrylamide gels after electrophoresis.

Keywords

Superoxide dismutase animal tissues erythrocyte human cancer tissues 

Abbreviations used

SOD

Superoxide dismutase

O2-

superoxide

KCN

potassium cyanide

NBT

nitroblue tetrazolium

GSH

glutathione

DTPA

diethylenetriaminepentaacetic acid

SDS

sodium dodecyl sulphate

Hb

haemoglobin

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References

  1. Ames, B. R., Cathcart, R., Schwiers, E. and Hochstein, P. (1981)Proc. Natl. Acad. Sci. USA,78, 6858.CrossRefPubMedPubMedCentralGoogle Scholar
  2. Azzi, A., Montecucco, C. and Richter, C. (1975)Biochem. Biophys. Res. Commun.,65, 597.CrossRefPubMedGoogle Scholar
  3. Beauchamp, C. and Fridovich, I. (1971)Anal. Biochem.,44, 276.CrossRefPubMedGoogle Scholar
  4. Beyer, W. F. Jr. and Fridovich, I. (1987)Anal. Biochem.,161, 559.CrossRefPubMedGoogle Scholar
  5. Bridges, S. M. and Salin, M. L. (1981)Plant Physiol.,68, 275.CrossRefPubMedPubMedCentralGoogle Scholar
  6. Crapo, J. D., McCord, J. M. and Fridovich, I. (1978)Methods Enzymol.,53, 382.CrossRefPubMedGoogle Scholar
  7. Davis, B. J. (1964)Ann. N. Y. Acad. Sci.,121, 404.CrossRefPubMedGoogle Scholar
  8. Flohé, L. and Ötting, F. (1984)Methods Enzymol.,105, 93.CrossRefPubMedGoogle Scholar
  9. Fridovich, I. (1974)Adv. Enzymol.,41, 35.PubMedGoogle Scholar
  10. Fridovich, I. (1983)Annu. Rev. Pharmacol. Toxicol.,23, 239.CrossRefPubMedGoogle Scholar
  11. Geller, B. L. and Winge, D. R. (1984)Methods Enzymol.,105, 105.CrossRefPubMedGoogle Scholar
  12. Gram, T. E., Okine, L. K. and Gram, R. A. (1986)Annu. Rev. Pharmacol. Toxicol.,26, 259.CrossRefPubMedGoogle Scholar
  13. Hartman, D. (1981)Proc. Natl. Acad. Sci. USA,78, 7124.CrossRefGoogle Scholar
  14. Leibovitz, B. E. and Siegel, B. V. (1980)J. Gerontol.,35, 45.CrossRefPubMedGoogle Scholar
  15. Lowry, O. H., Rosebrough, N. J., Farr, A. L. and Randall, R. J. (1951)J. Biol. Chem.,193, 265.PubMedGoogle Scholar
  16. Marklund, S. L. and Marklund, G. (1974)Eur. J. Biochem.,47, 469.CrossRefPubMedGoogle Scholar
  17. McCord, J. M. and Fridovich, I. (1969)J. Biol. Chem.,244, 6049.PubMedGoogle Scholar
  18. Misra, H. P. and Fridovich, I. (1972)J. Biol. Chem.,247, 3170.PubMedGoogle Scholar
  19. Nandi, A. and Chatterjee, I. B. (1987)J. Biosci.,11, 435.CrossRefGoogle Scholar
  20. Nishikimi, M. (1975)Biochem. Biophys. Res. Commun.,63, 463.CrossRefPubMedGoogle Scholar
  21. Oberly, L. W. and Buettner, G. R. (1979)Cancer Res.,39, 1141.Google Scholar
  22. Oberly, L. W. and Spitz, D. R. (1984)Methods Enzymol.,105, 457.CrossRefGoogle Scholar
  23. Richterich, R. (1969)Clinical chemistry (Basel, Switzerland, New York: S. Karger).Google Scholar
  24. Rigo, A., Viglino, P. and Rotilio, G. (1975)Anal Biochem.,68, 1.CrossRefPubMedGoogle Scholar
  25. Peeters-Jorris, C., Vandevoorde, A. M. and Bandhuin, P. (1975)Biochem. J.,150, 31.CrossRefGoogle Scholar
  26. Steinhagen-Thiessen, E., Reznick, A., Ringe, J. D. and Gershon, D. (1986) inSuperoxide and superoxide dismutase in chemistry, biology and medicine (ed. G. Rotilio) (Amsterdam: Elsevier) p. 464.Google Scholar
  27. Sykes, J. A., McCormack, F. X. Jr. and O’Brien, J. J. (1978)Cancer Res.,38, 2759.PubMedGoogle Scholar
  28. Takada, Y., Noguchi, T., Okabe, T. and Kajiyama, M. (1982)Cancer Res.,42, 4233.PubMedGoogle Scholar
  29. Troll, W. and Weisner, R. (1985)Annu. Rev. Pharmacol Toxicol.,25, 509.CrossRefPubMedGoogle Scholar
  30. Westman, N. G. and Marklund, S. L. (1981)Cancer Res.,41, 2962.PubMedGoogle Scholar

Copyright information

© Indian Academy of Sciences 1988

Authors and Affiliations

  • Anuradha Nandi
    • 1
  • I. B. Chatterjee
    • 1
  1. 1.Department of BiochemistryUniversity College of ScienceCalcuttaIndia

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