I want to do antioxidant enzymatic assay of plant sample. For that will i have to do the assay of pure enzymes also (standard) for each enzymatic assay? How to calculate the enzyme concentration in this assay plz suggest me
Thanking you.
Dear Anupam Apoorva,
It is expected that your plant sample will have more than one enzyme. Therefore, it will incorrect to calculate the enzyme concentration in your sample. What you can do is to know what kind of enzymes this plant may have and to run enzymatic reactions for each of these enzymes (standard) and compare the enzymatic activity of your plant sample to those standards. In a next step, you may be able to purify the enzymes from your plant and run enzymatic reaction for each of the pure enzyme and compare it to known standard enzymes.
Hoping this will be helpful,
Rafik
Dear Anupama,
I presume that you are wanting to to explore the presence of common antioxidant enzyme activities in your plant extract. Your plant samples can be analyzed for general antioxidant enzymes such as catalase, SOD, glutathione peroxidase and so on by following standard enzyme assay methods (described in the series, Methods in Enzymology), using different protein concentrations of your sample. In order to ascertain that you are properly determining a given enzymic activity in your sample you can perform an assay of the commercial standard of the particular enzyme in question. You can find assay methods online. Sigma Aldrich's website is one such source.
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