Assay Method Information

Assay Name:  The enzymatic inhibition assay
Description:  The enzymatic inhibition assay was carried out with a final concentration of 100 nM recombinant SARS-CoV-2 Mpro and 20 μM FRET substrate. For the determination of IC50 values, the enzyme was pre-incubated with various concentrations of each compound for 10 min, and then the reaction was initiated by adding the FRET substrate and monitored at 405 nm in the kinetic mode for 10 min on a CLARIOstar microplate reader (BMG Labtech). The initial velocities were calculated from the first 1 min of each reaction curve. The IC50 values were calculated using a dose-response model in GraphPad Prism 8.0 software. All experiments were performed in triplicate, and the values are presented as mean ± standard deviation (SD).
Affinity data for this assay
 

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