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Assay Method Information

Assay Name:  Antimalarial Testing In Vitro (IC50) and Measurement of Inhibition Constant (Ki)
Description:  The concentration of inhibitor that inhibited 50% of the parasite growth (IC50) was determined from the sigmoidal curve obtained by plotting the percentages of [3H]-hypoxanthine incorporation against drug concentrations. The activity of pfDHFR-TS was determined spectrophotometrically using a UV-Vis spectrophotometer. The Ki values of the inhibitors for the wild-type and mutant enzymes were determined by fitting to the equation IC50 = Ki(1 + ([S]/Km)), where IC50 is the concentration of inhibitor that inhibits 50% of the enzyme activity under the standard assay condition and Km is the Michaelis constant for the substrate H2folate.
Affinity data for this assay
 

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Last update November 1, 2007
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