| Assay Method Information | |
| | MERS-CoV MPro Enzyme inhibition IC50 determination |
| Description: | The inhibition of the MERS-CoV-2 MPro Enzyme was measured by an analogous method to that of Mesecar et al.20 The fluorescence of the cleaved FAM peptide (excitation 480 nm/emission 520 nm) is measured using a fluorescence intensity protocol on a Pherastar FX (BMG).The assay reaction buffer contained 20 mM HEPES (pH 7.3), 50 mM NaCl, 1 mM TCEP, 0.1 mg/ml BSA and 0.01% Triton x-100. Compounds were pre-incubated with 50 nM MERS 3CL protease for 15 minutes at 23° C. Enzyme reactions were initiated with the addition of 550 nM (5-FAM)-GVLQSGLV-K(Dabcyl)-K-NH2 substrate and allowed to proceed for 60 minutes at 23 °C. Percent inhibition or activity was calculated based on control wells containing no compound (0% inhibition/100% activity). IC5o values were generated using a 4-parameter hill slope fit model using Vault software (CDD). |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |