| Assay Method Information | |
| | Cap-Dependent Endonuclease (CEN) Inhibitory Activity Assay |
| Description: | 2.5 μL of enzymatic reaction mixture (composition: 53 mM Tris hydrochloride (pH 7.8), 1 mM MgCl2, 1.25 mM dithiothreitol, 80 mM NaCl, 12.5% glycerol, 0.15 μL enzyme solution) was dispensed in a 384-well plate made of polypropylene. Then, 0.5 μL of DMSO was added to 0.5 μL of the test compound solution serially diluted with DMSO, the positive control (PC) and the negative control (NC), and mixed thoroughly. Then 2 μL of substrate solution (1.4 nM substrate RNA, 0.05% Tween20) was added, and the reaction was initiated. The reaction mixture was incubated at room temperature for 60 minutes, and then 1 μL of the reaction mixture was added to 10 μL of high-purity formamide solution (containing GeneScan120LizSize Standard as sizing marker, manufactured by Applied Biosystem(ABI)) to terminate the reaction. The reaction of NC was stopped in advance by adding EDTA (4.5 mM) before the start of the reaction (all labeled concentrations were the final concentrations). |
| Affinity data for this assay | |
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