Groups of data (e.g. wells of a plates) are normalized relative to two subsets (e.g. some wells within that plate, e.g. some negative control wells and some positive controls wells). For each group the mean (or median) is calculated for the two selected subsets. Based on these estimates, all data points of each group are normalized by applying this formula:
x.npi = (mean(x[subset{positive}]) - x) / (mean(x[subset{positive}]) - mean(x[subset{negative}])) * 100
or
x.npi = (median(x[subset{positive}]) - x) / (median(x[subset{positive}]) - median(x[subset{negative}]))
* 100
Therfore x.npi values resemble the "percentage of inhibition", where the positive subset is defined as 0% and the negative subset as 100%. By swapping the two subsets the definition will be "percentage of activation" and negatives will be set to 0% and positives to 100%.
Literature: Malo et al., Nat. Biotechnol. 24, 167-179 (2006)
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