I am using the 19160 SOD Assay Kit-WST (Sigmma) for my SOD experiment between young flies and old flies. I found that young flies has % inhibition of 60, while old flies has % inhibition of 30. Using the following equation: SOD activity (inhibition rate %) = {[(Ablank 1 - Ablank 3) – (Asample - Ablank 2)]/ (Ablank 1 - Ablank 3)} x 100
Here is my question, is that mean old flies has more SOD because of the low conversion rate of WST-1 to WST-1 formazan. Please help.
For this experiment, I measured the Endpoint, not kinetic. I want to compare the SOD activity between young and old flies. Also, is it possible to report my results as Units/ml Enzyme? If so, what equation should I use?
Thank you so much for your time.
Hi Bacha,
Thank you for the answer. Do you think it is possible to calculate the units/ml enzyme using the equation below? Thanks
CALCULATIONS
Percent Inhibition:
SOD activity (inhibition rate %) = {[(Ablank 1 - Ablank 3) – (Asample - Ablank 2)]/ (Ablank 1 - Ablank 3)} x 100
Units/ml Enzyme:
[(Percent Inhibition)(DF)] / [(50%)(0.10)]
DF = Dilution Factor
50% = Inhibition of the rate of cytochrome c reduction per the unit definition
0.10 = Volume (in milliliters) of enzyme used in each test
Dear Hun, first you should correct the SOD activity per mg of proteins present in samples.
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