| Assay Method Information | |
| | Enzymatic Assay |
| Description: | The main protease (Mpro) enzymatic assays were carried out exact as previously described in pH 6.5 reaction buffer containing 20 mM HEPES pH 6.5, 120 mM NaCl, 0.4 mM EDTA, 20% glycerol and 4 mM DTT.3. The SARS-CoV-2 papain-like protease (PLpro) enzymatic assays were carried out as follows: the assay was assembled in 96-well plates with 100 μl of 200 nM PLPro protein in PLPro reaction buffer (50 mM HEPES, pH7.5, 0.01% triton-100 and 5 mM DTT). Then 1 μl testing compound at various concentrations was added to each well and incubated at 30° C. for 30 min. The enzymatic reaction was initiated by adding 1 μl of 1 mM FRET substrate (the final substrate concentration is 10 μM). The reaction was monitored in a Cytation 5 image reader with filters for excitation at 360/40 nm and emission at 460/40 nm at 30° C. for 1 hr. The initial velocity of the enzymatic reaction with and without testing compounds was calculated by linear regression for the first 15 min of the kinetic progress curve. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |