| Assay Method Information | |
| | hDHODH Inhibition Assay |
| Description: | Inhibitory activity was assessed by monitoring the reduction of 2,6-dichloroindophenol (DCIP), which is associated with the oxidation of dihydroorotate as catalyzed by the DHODH enzyme. The enzyme was preincubated for five minutes at 37° C. in Tris-buffer solution (pH 8.0), with coenzyme Q10 (100 μM), with the compounds to be tested used at different concentrations (final DMSO concentration 0.1% v/v), with DCIP (50 μM). The reaction was initiated by the addition of DHO (500 μM), and the reduction was monitored at λ=650 nm. The initial rate was measured in the first five minutes (ε=10400 M−1 cm−1), and an IC50 value was calculated,31 using GraphPad Prism 7 software. Values are means±SE of three independent experiments. |
| Affinity data for this assay | |
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