I need to analyse the oxidative stress of my mice tissue samples. So I need standard simple methods to perform it. I need to analyse the following enzymes, viz, Superoxide dismutase, Glutathione peroxidase, Catalase and Lipid peroxidase. So please anyone can anyone attach some spectrophotometric methods to do these assays?
Kits available from Trevigen Inc. are robust and provides much reproducible results. We have a long experience using these kits in various tissue types. We may please refer to the following publications from my group:
Article Occult hepatitis C virus elicits mitochondrial oxidative str...
Article Role and clinical significance of lymphocyte mitochondrial d...
You can use expensive kits or spectrophotometric methotd
SOD:
Misra H.P, Fridovich J.: The role of superoxide anion in the autooxidation of epinephrine and simple assay for superoxide dismutase. J. Biol. Chem. 1972; 247(31): 70-3175
CAT:
Beers R., Sizer T.: Spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J. Biol. Chem. 1952; 195: 133-140
GPX:
Paglia DE, Valentine WN.Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase.J Lab Clin Med. 1967 Jul;70(1):158-69.
you will find in attachment a detailed protocol for the determination of 3 enzymes; SOD, GPX and CAT. but this protocol is used for plants.
You can use Ransel and Ransod kits for GPx and SOD.
MDA and CAT also assay with bichemical methods.
You can also see this article published in Nature protocols (2010) (file attached)
The techniques used : Western blots, activity gels and activity assays for SOD, CAT and GPX activities
Standard methods for several antioxidant parameters with description used in our laboratory.
https://www.researchgate.net/publication/340429895_Supplementary_material_manuscriptv7docx
I you have any questions freely write,
Marko
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