I carried out SOD, GST and Catalase activity from the different plants of family Asteraceae in normal condition no stress nd all.
Now i want to know should i use a standard(i.e Enzyme SOD and Catalase) for comparison or for the expression of results or i can express the results without any standard just in fresh weight or protein.
Antioxidant enzyme activities?
The antioxidant enzymes activities: SOD, Peroxidase and Catalase could be realized and expressed as U/mg of protein (Specific Activity) or U/ml of extract without comparison with standard enzymes. You may have to do comparison with results discussed in literature.
However, you have to take into account that the enzymes extracted from plants must be done with optimized conditions to obtain the best yield of native activities.
Yet I want to advise you to test oxidants enzymes with the standard spectro-photometric methodusing the free plant extract. That test will give you each specific enzyme activity (U/ml or U/mg).
On another hand I advise you to test enzymes with the zymography technique, that method may inform you about the isoenzymes of each enzyme.
In attached file I send you a manuscript that discusses these methods tested in our laboratoryDr Imen Hajji Sfaxi, Dr Feten belhadj (Materiel and methods and native PAGE zymogram)
Good work
Best regards
Antioxidant enzyme activities?
The antioxidant enzymes activities: SOD, Peroxidase and Catalase could be realized and expressed as U/mg of protein (Specific Activity) or U/ml of extract without comparison with standard enzymes. You may have to do comparison with results discussed in literature.
However, you have to take into account that the enzymes extracted from plants must be done with optimized conditions to obtain the best yield of native activities.
Yet I want to advise you to test oxidants enzymes with the standard spectro-photometric methodusing the free plant extract. That test will give you each specific enzyme activity (U/ml or U/mg).
On another hand I advise you to test enzymes with the zymography technique, that method may inform you about the isoenzymes of each enzyme.
In attached file I send you a manuscript that discusses these methods tested in our laboratoryDr Imen Hajji Sfaxi, Dr Feten belhadj (Materiel and methods and native PAGE zymogram)
Good work
Best regards
Sir can you send calculations of Catalase and GST am finding a little bit confusion in calculations.
There is no need for any stander in the determination of these antioxidant enzymes
Hello! You don´t need to use a standard. You can expressed as U/mg of protein or U/mg of fresh weight.
Best regards!
Hello Lalita you donot need to used standards for these enzymes you can express the enzyme activity as unit enzyme per mg protein see what species of your plant family has the lower activity and consider this activity as 100% the calculate for other species and report it relative to this as for example 120%,140%etc also compare your enzymes activity with other reported in literature for other plant family species.For assay of these enzymes you can used commercially available kits from Sigma chemicals or others also you can see my RG for these enzymes from other sources good luck
No need to use standard. Express you activities as units/mg protein
These is no need to use standard. Express you activities as units/mg protein.
You can express your activities as units/mg protein. There is no ned to compare with standard enzymes like catalase/ SOD etc.
You can express your activities as units/mg protein. There is no need to compare with standard enzymes like catalase/ SOD etc.
Antioxidant enzyme activity are several enzyme systems that catalyze reactions to neutralize free radicals and reactive oxygen species. These enzymes include:
- superoxide dismutase
- glutathione peroxidise
- glutathione reductase
- catalases
These form the body’s endogenous defence mechanisms to help protect against free radical-induced cell damage. The antioxidant enzymes – glutathioneperoxidase, catalase, and superoxide dismutase (SOD) – metabolize oxidative toxic intermediates.
These enzymes also require co-factors such as selenium, iron, copper, zinc, and manganese for optimum catalytic activity. It has been suggested that an inadequate dietary intake of these trace minerals may compromise the effectiveness of these antioxidant defense mechanisms. The consumption and absorption of these important trace minerals may decrease with aging.
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