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| | HCV RNA-Dependent RNA Polymerase Assay (IC50) and Replicon Cell-Based ELISA Assay (EC50) |
| Description: | Primer-dependent assays were performed using the homopolymeric template/primer. Compounds, polymerase and template RNA were incubated at RT for 25 min before the addition of NTPs. Alternatively, compounds were added to the preformed polymerase/template complex and incubated at RT for 10 min before the addition of 10 uM UTP, 2 uCi 3H-UTP. Elongation proceeded for 1 hr at RT and the activity was measured as acid insoluble radioactivity. In the single turnover experiments, elongation reaction was started by addition of nucleotides and 50 ng/ul heparin. IC50 values were calculated using three parameters logistic equation and inhibition data were fitted by Kaleidagraph software. The effect of compounds (EC50) on viral replication was monitored by Cell-ELISA. |
| Affinity data for this assay | |
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