| Assay Method Information | |
| | DHODH Inhibition Assay |
| Description: | The assays were carried out by using a colorimetric DCIP method, which uses the colorimetric reagent 2, 6-dichlorophenolindophenol as the final electron acceptor. DCIP reduction is stoichiometrically equivalent to oxidation of dihydroorotate. Changes in absorbance were quantified in triplicate on a plate reader and data were analyzed to measure the reduction of DCIP as a decrease in absorbance at 595 nm. Microsoft Excel was used to plot and analyze the data to determine kinetic parameters. Initial reaction rates were calculated by least-squares linear regression. Lineweaver-Burke plots suggested initial guesses for the kinetic parameters and the type of inhibition to be fitted. Kic values were obtained for compounds displaying competitive inhibition. |
| Affinity data for this assay | |
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