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Superoxide dismutase micro assay in biological material.
Hoppe-Seyler'S Zeitschrift Fur Physiologische Chemie
|June 1, 1975
Summary
A new micro assay accurately measures superoxide dismutase (SOD) activity in small biological samples. This method uses gel electrophoresis and colorimetry, proving effective even in complex biological mixtures.
Area of Science:
- Biochemistry
- Enzymology
Background:
- Superoxide dismutase (SOD) is a crucial antioxidant enzyme.
- Accurate measurement of SOD activity is vital for biological research.
- Existing assays may require larger sample volumes or are less sensitive.
Purpose of the Study:
- To develop a rapid and convenient micro assay for determining superoxide dismutase activity.
- To validate the assay's performance using limited biological material.
- To establish a reproducible method for quantifying SOD levels.
Main Methods:
- Utilized formazan derivative color formation from nitroblue tetrazolium reduction by superoxide radicals (O2•−).
- Integrated analytical polyacrylamide gel electrophoresis (PAGE) with the colorimetric reaction.
- Employed a gel-scanning apparatus to quantify enzyme activity based on unstained gel regions.
Main Results:
- Demonstrated a linear relationship between soluble SOD reactivity in gels and enzyme concentration.
- Established a reproducible calibration curve where integrated scanning signal correlated logarithmically with SOD activity (10⁻¹² to 7 × 10⁻¹¹ mol).
- Successfully detected added SOD in crude liver and brain homogenates, showing no inhibition by biological components.
Conclusions:
- The developed micro assay is effective for rapid and sensitive SOD activity determination.
- The assay is robust and applicable to various biological samples, including crude homogenates.
- This method offers a reliable tool for biochemical and biomedical research requiring SOD quantification.