| Assay Method Information | |
| | 3CL protease inhibition assay |
| Description: | The 3CL protease inhibition assay was conducted in 384-well plates (Corning 3702). The substance solution (10 mM dimethyl sulfoxide [DMSO] solution) was diluted to 250 µmol/L stepwise with a 3-fold dilution with DMSO. Finally, the solutions were mixed with 20 mmol/L Tris-HCl (pH 7.5) as a compound solution. Ten microliters of compound solution was added manually to each well, then 5 µL of 16 µM substrate in inhibition buffer (2 mM EDTA, 20 mM DTT, 0.02% BSA, and 20 mM Tris-HCl, pH 7.5) was added. The reaction was initiated by adding 5 µL of 12 nM 3CL protease in inhibition buffer and incubated at room temperature for 3 h. The following operations were the same as those described in the biological screening. |
| Affinity data for this assay | |
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