i have some data about a recombinant enzyme and i want to calculate my enzyme unite.
i have used the ellman method for enzyme activity test.
ODfirst minite= 0.35
ODseconde minite=0.8
reaction volume=0.2 ml (standard 96-plait, high 10.63 mm)
substrate concentration=1 mM
enzyme concentration= 0.5 mg/ml
You need an additional piece of information to convert the OD to the product concentration. It could either be the extinction coefficient change of the product or, preferably, a standard curve of product concentration versus absorbance under identical assay conditions to the ones you used.
In addition, you need to find out whether the reaction rate was constant during the 2 minute measurement. In other words, were you measuring the initial rate of the reaction. It would also help to know what the background absorbance was of the same reaction omitting only the enzyme. This should be subtracted from the enzyme-catalyzed reaction's absorbance.
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