Or should it be calculated using the linear equation Or should the concentration be converted in to log values before plotting a graph and the equation obtained should be used for calculation? Which one is ideal? What could be the reason, if there is a mismatch between the dose-response graph and the reported IC50 value? Does the method used for calculation affect the IC50 value in an enzyme inhibition assay? What is the ideal method for calculation of IC50 value?
The IC50's won't match If you are performing enzyme inhibition assays using a different reaction condition [ like substrate, buffers, co-factors, temperature etc.,] as that of reported for the determination of IC50 of the compound of interest. In such case you can go for the Ki value for that inhibitor which would not vary for an inhibitor even if there exists differences in assay method/conditions.
Did you determined the confidence interval of your IC50, to ensure that the mismatch is not due to lack of precision or something like that?
The calculation method may influence it, but also if you're concentrations are not low and high enough to determine the Emin and Emax values, the IC50 will not be determined correctly.
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