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| | Wild Type SARS-CoV-2 Mpro Inhibition Assay |
| Description: | Mpro activity was measured according to the manufacturer's protocol (BPS Biosciences, cat #78042-2) in 384-well plates. The included recombinant un-tagged Mpro (referred to as 3CL protease in the kit) was tested for ability to cleave a FRET peptide substrate (DABCYL-KTSAVLQSGFRKME-EDANS) to generate a fluorescent product (SGFRKME-EDANS) over time. The final volume is 25 ml in indicated assay buffer, with final Mpro and substrate concentrations as 17 nM and 40 uM respectively. Fluorescent intensities were monitored once every 5 minutes after adding substrate into the rest of reaction mixture. Enzyme activity was represented using the initial linear slope of the kinetic curve. Compounds were first serially diluted in DMSO as 1000 working stock, then diluted in assay buffer before adding to reaction mixture following manufacture's protocol. 100 mM GC376 was used as 100% inhibition control. For no inhibition control, no compound was added. Blank wells contained buffer, substrate but no Mpro. Final 0.1% DMSO was included as vehicle no inhibition control. |
| Affinity data for this assay | |
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