| Assay Method Information | |
| | Inhibitory Activity Against Mpro |
| Description: | An objective of this experiment is to detect the in vitro inhibitory activity against Mpro of the compound of the present disclosure.Experimental materials:GST-AVLQ-3Clpro-GP-6His (CP, Lot. No. CP20200526001);MCA-AVLQSGFR-Lys(Dnp)-Lys-NH2 (GL, Customized); and384-well plate (Perkin Elmer, Cat. No. 6007279).Experimental steps and methods:1. Prepare a 1× buffer solution (modified Tris buffer solution).2. Transfer a to-be-tested compound to the assay plate with Echo's 100% DMSO solution to a final DMSO content of 0.5%.3. Prepare a mixed protein solution: Add GST-AVLQ-3Clpro-GP-6His in a 1× assay buffer solution to support the mixed protein solution.4. Prepare a mixed substrate solution: Add the polypeptide (MCA-AVLQSGFR-Lys(Dnp)-Lys-NH2) in a 1× assay buffer solution to obtain the substrate solution.5. Add 10 μL of 2× mixed protein solution in the assay plate, centrifuge for 30s, shake for 30s, and incubate the solution at room temperature for 15 min.6. Add 10 μL of 2× mixed substrate solution in the assay plate, centrifuge for 30, and shake for 30s.7. Read the data dynamically on Synergy (Ex320/Em405). |
| Affinity data for this assay | |
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