| Assay Method Information | |
| | Probe Displacement Assay |
| Description: | The probe displacement assay is conducted as follows: In a 384-well plate, test compound along with recombinantly expressed His-tagged protein corresponding to amino acids 556-871 of human TYK2 (sequence shown below) at 5 nM, 33 nM probe (BB, preparation described below) and 0.33 mg/mL Yttrium oxide (YOx) His-capture SPA beads (PerkinElmer, RPNQ0276) in IX PBS containing 10 mM MgCl2, 0.1 mg/mL BSA, 0.1 mM TCEP, 0.005% Tween-20, and 1% DMSO (final concentration) were incubated for 10 hours at room temperature. The amount of radiolabeled probe bound to TYK2 was then quantified using a ViewLux Microplate Imager, and the fraction bound by the test compound calculated by comparison to wells containing scale references for 0% and 100% probe displacement. The IC50 value is defined as the concentration of test compound required to inhibit radiolabeled probe binding by 50%. |
| Affinity data for this assay | |
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