| Assay Method Information | |
| | DENV2 Protease Assay |
| Description: | To measure the inhibitory activities of the compounds, the DENV2 NS2B-NS3 protease activities were measured in the presence of the compounds using Bz-Nle-Lys-Arg-Arg-AMC as a substrate. The enzyme concentration in the assay was 125 nM, and the substrate concentration was 20 µM in 10 mM Tris-HCl pH 8.5, 20% glycerol, and 1 mM CHAPS. The inhibitor concentrations were 100 µM for the first round and between 3.125 and 400 µM for the compounds 16 and 19 in the second round. Before adding the substrate, the enzyme and the inhibitors were incubated at 37 °C for 15 min. All reactions were performed in 96-well plates with a final volume of 50 µL per each wells using the FLx800 fluorescence microplate reader (excitation = 380 nm; emission = 460 nm). |
| Affinity data for this assay | |
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