| Assay Method Information | |
| | ADA Inhibition Assay |
| Description: | The target compounds were screened against calf intestine ADA in vitro by monitoring the rate of hydrolyzing adenosine into inosine spectrophotometrically. While holding the enzyme and inhibitor concentration constant, the substrate concentration was varied to obtain the kinetic data. The substrate concentration in each assay was measured using up to five different concentrations, ranging from 20 to 60 uM. This was repeated with a different inhibitor concentration. The enzyme concentration was kept constant in all assays. Lineweaver-Burk plots were constructed for each assay and the Ki was calculated using Graph Pad Prism software. Assays were performed in duplicate and in some cases triplicate. |
| Affinity data for this assay | |
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