I need the procedures to measure the antioxidant levels and activities such as Superoxide dismutase, catalase, Lipid peroxide, Reduced glutathione, glutathione peroxidase and glutathione-S- transferase in the rat tissue samples such as liver, kidney and pancreas.
For measuring the effect of free radicals on the tissue and subsequent damage, u can go for lipid peroxidation assay to measure the inhibition factor......or else if u just want to do the antioxidant assay then u can go for FRAP assay , where u can take the sample that is around 100 µl (Plasma/ Milk/ Urine/ Feed Extract etc.) and mix it with 3 ml of working FRAP reagent and measure the absorbance at 593 nm.
Refer this link...its a free article...
http://nopr.niscair.res.in/bitstream/123456789/14404/1/IJNPR%203(2)%20228-231.pdf
for detailed protocol of frap assay, refer:
http://www.scribd.com/doc/4940425/FRAP-Assay
http://www.scribd.com/doc/76649053/FRAP-Protocol
what type of antioxidant? is it enzymatic antioxidants such as catalase, superoxide dismutase, glutathione peroxidase, glutathione S-transferase, glutathione reductase activities OR non-enzymatic antioxidants such as reduced glutathione, ascorbic acid....
I have not done any research on rat, but for poultry I have determined the activity of antioxidant enzymes, TBARS and total antioxidant capacity in meat that could be practical for rat tissues. You can see more information in these papers.
Article Effect of dietary turmeric on antioxidant properties of thig...
Article Short communication. Dietary supplementation effects of zinc...
Here is a review that compare different methods for assessing antioxidant capacity in human serum.
Thank you all, I found all your information and articles useful. I am currently working on determination of antioxidants enzymes and immunology parameters in chickens
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