| Assay Method Information | |
| | TYK2 JH2 Domain Binding Assay |
| Description: | Assays were performed in 384-well plates with a final assay volume of 20 μL containing copper-polyvinyltoluene scintillation proximity assay (SPA) beads (PerkinElmer Life Sciences, catalogue no. RPNQ0095) at 80 g/ml, H3 probe (20 nM), the N-terminal His-tagged TYK2 pseudokinase domain (2.5 nM, purified by Pharmaron), and test compounds in assay buffer (50 mM HEPES, pH 7.5, 100 g ml-1 BSA, 5% DMSO). After incubating at room temperature for 30 min, the inhibition was calculated by the displacement of [3H] 3 binding as determined by scintillation counting. Dose-response curves were generated to determine the concentration required to inhibit H3 probe binding by 50% (IC50). |
| Affinity data for this assay | |
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