| Assay Method Information | |
| | High-Throughput Screening and IC50 Determinations |
| Description: | The screening assay was performed using 384-well microtiter plate first spotted with test compounds. Then Ape1 in reaction buffer was added to each well. The reaction was initiated through the addition of substrate. After incubation, the final fluorescence was measured using a FLUOstar Optima plate reader (BMG Labtech GmbH, Offenburg, Germany) at excitation and emission wavelengths of 485 and 520 nm, respectively. IC50 analyses were carried out on the best hits by first plating 10-fold dilutions of each inhibitor onto 384-well plates. Using the initial rate values, percentage activity was calculated for each sample relative to a negative (DMSO only) control. The data were fitted to a sigmoidal dose-response model using Prism software (GraphPad Software, San Diego, CA). |
| Affinity data for this assay | |
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