I am conducting experiments about enzyme kinetics. And I am confused about data normalization. For example, Uv absorbance of the enzymic reaction is usually used to monitor the process. I have draw a Abs-Time curve about a reaction. How can I Normalized the curve? Abs(t=x)/Abs(t=0)? Or Abs(t=x)-Abs(t=0)?
Not sure about the uv. You may normalise the data but maybe from the absorbance. Maybe your first option. It will be like densitometry on a western blot. You pick the data from the y axis
You do not need to normalize the data. The analysis you should do is to measure the slope of the progress curve (delta absorbance/delta time) at the beginning when it is steepest. This slope is directly proportional to the initial rate of the reaction.
The next step is to convert the slope to the rate of product formation (or substrate consumption, depending on what you are measuring by absorbance). This requires knowing the extinction coefficient change that occurs upon conversion of substrate to product, which can be measured by making standard curves of product and substrate at various concentrations.
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