| Assay Method Information | |
| | TYK2 JH12 Domain Binding Assay |
| Description: | Binding constants for compounds of the present invention against the JH2 domain were determined by the following protocol for a KINOMVEscan® assay (DiscoveRx). A fusion protein of a partial length construct of human TYK2 (JH12domain-pseudokinase) (amino acids G556 to D888 based on reference sequence NP_003322.3) and the DNA binding domain of NFkB was expressed in transiently transfected HEK293 cells. From these HEK 293 cells, extracts were prepared in M-PER extraction buffer (Pierce) in the presence of Protease Inhibitor Cocktail Complete (Roche) and Phosphatase Inhibitor Cocktail Set II (Merck) per manufacturers' instructions. The TYK2 (JH2domain-pseudokinase) fusion protein was labeled with a chimeric double-stranded DNA tag containing the NFkB binding site (5′-GGGAATTCCC-3′) fused to an amplicon for qPCR readout, which was added directly to the expression extract (the final concentration of DNA-tag in the binding reaction is 0.1 nM). |
| Affinity data for this assay | |
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