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| | fluorescence resonance energy transfer (FRET)-based substrate cleavage assay |
| Description: | The respective human coronavirus Mpro in assay buffer (20 mM Tris-HCl, pH 7.3, 100 mM NaCl, 1 mM EDTA, 5 mM TCEP) and 0.1% BSA was added to assay-ready plates containing compound. The enzymatic reaction was then immediately initiated with the addition of 5 μl substrate in assay buffer. Final concentrations of respective protease and substrate are shown in the table below. Initial rates were measured by following the fluorescence of the cleaved substrate using a Spectramax (Molecular Devices) fluorescence plate reader in the kinetic format. |
| Affinity data for this assay | |
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