Can anybody help me to calculate the SOD activity using the method of Dhindsa et al., 1981?
I am not familiar with this method - Dhindsa et al., 1981! Can you give me a complete citation? Why don't you use the Original Cytochrome C method by McCord and Fridovich?
There is also the "Mistra and Fridovich, 1972" method which involves the auto-oxidation of adrenaline by SOD.
This chapter of Methods in Enzymology contains the standard assay for SOD activity, and explains how it is calculated from competition experiments. I hope it helps.
http://www.sciencedirect.com.ezproxy3.lhl.uab.edu/science/article/pii/S0076687978530449
Hello Dhiraj
You can calculate SOD Activity as follows:
(The reaction mix. is to be prepared in 50mM Phosphate buffer-pH 7.8)
The 3.5 ml reaction mix contains
6 microM NBT
13 mMMethionine
0.1 mM EDTA
13 microM Riboflavin
0.05M Na2CO3
0.5ml Enzyme extract
O.D is read at 560nm (Decrease in OD wrt Control (without Enzyme))
OD of Blank - OD of C(Light - dark) = x = I unit of enzyme
OD of C/2
The EU in Reaction Mixture / enzyme extract = OD/x
Try this and tell me if ur problem is sorted.
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