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I've just started with inhibition studies on the metabolism of THC and
before starting the experiments, I would like to know some very general thin=
1.Are there any rules or a "general strategy" how to perform the Km and Ki
determinations, especially how I find the concentration ranges of
substrate/inhibitor to be used? Of course the concentration of the substrate
and/or the inhibitor are extremely important and they have to be considered
very carefully. But in all the literature I've looked, they used ranges of
substrate/inhibitor without any explanations, so I'm not sure if their
ranges are the result of several pre-experiments, or if the author's simply
took some basics or generalities of enzyme kineticsinto consideration, which
pre-set these ranges more or less inevitably.
2. In most literature, as unit of velocity they used =B5M per min, but the
appropiate incubation times were mostly 10 minutes or longer. So, in order
to reach the "one-minute-unit" of velocity (in order to draw
Lineweaver-Burk-plots), is it possible to devide the absolute found value
(e.g. 10 min) of velocity by 10?
Thank you very much in advance for helping me to find the "basics"!
University of Saarland
Institute of Legal Medicine, Toxicology
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