| Assay Method Information | |
| | SARS-CoV-2 Mpro Protease Enzyme Assay |
| Description: | It was performed in 20 mM Tris, 50 mM NaCl and 0.1 mM EDTA (Merck KGaA, Darmstadt, Germany), pH 7.5 at room temperature. Compounds were transferred with Echo 550 non-contact dispenser (Labcyte, Inc., USA) to a Corning 3575 non-binding 384 well assay plates. Mpro (75 nM final concentration), was added to the assay plate using a 16-channel pipette (Integra ViaFlo, BergmanLabora AB, Sweden), and shaken for 15 minutes at 1500 rpm in an Eppendorf Mixmate. After a pulse centrifugation, the Mpro fluorogenic substrate (stock solution at 5 mM in DMSO) was added to the assay plate to a final concentration of 10 μM, thus contributing with 0.2% DMSO in final assay, with a Labcyte ECHO 550 non-contact dispenser. After 10 minutes incubation and a pulse centrifugation, fluorescence was measured in a PerkinElmer Envision plate reader at ambient temperature using kinetic mode and with excitation at 340 nm and emission at 490 nm. |
| Affinity data for this assay | |
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