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Analysis of superoxide dismutase activity
1University of Colorado Health Science Center, Denver, Colorado, USA.
Current Protocols in Toxicology
|October 10, 2012
Summary
Measuring superoxide dismutase (SOD) activity is difficult due to substrate instability. This study details methods to indirectly assay SOD activity by monitoring competition with indicator molecules or using activity stains on gels.
Area of Science:
- Biochemistry
- Enzymology
- Oxidative Stress
Background:
- Superoxide dismutases (SODs) are crucial enzymes that manage cellular hydrogen peroxide levels.
- Assaying SOD activity is challenging due to the instability of its substrate, superoxide, at physiological pH.
Purpose of the Study:
- To describe reliable methods for measuring total SOD activity and specific isoforms like CuZn-SOD and MnSOD.
- To overcome the challenges associated with direct substrate measurement.
Main Methods:
- Indirect activity measurement through competition assays between SOD and indicator molecules reacting with superoxide.
- Direct activity assessment using activity stains on thin-film agarose or native polyacrylamide gels.
Main Results:
- The described indirect method allows for the quantification of total SOD activity.
- Specific SOD isoforms (CuZn-SOD, MnSOD) can be differentiated and their activities measured.
- Activity staining provides an alternative visualization and quantification method.
Conclusions:
- Established indirect and gel-based activity staining methods enable robust measurement of SOD enzyme activity.
- These techniques are valuable for studying oxidative stress and the role of SOD enzymes in biological systems.
