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For the FP competition assay, the unlabelled compounds (from 50-mM DMSO stocks) were first dispensed in duplicate into wells of a Proxiplate 384 F Plus plate using an Echo 550 acoustic liquid handler to give a 13-point concentration range from 0.2 to 500 M

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Subsequently, we added DTC, as a Pd scavenger 16 , and a fresh amount of DSF followed by adjustment of the reaction to pH 7

To do so, measured M concentration of target metabolite in the erythrocyte fraction was divided on concentration factor after gradient