| Assay Method Information | |
| | In Vitro Activity Assay of JAK1, JAK2, JAK3, TYK2 Kinases |
| Description: | JAK1(h) was incubated with 20 mM Tris/HCl pH 7.5, 0.2 mM EDTA, 500 μM MGEEPLYWSFPAKKK, 10 mM magnesium acetate and [γ-33P]-ATP (activity and concentration were determined as required). The reaction was started by adding the Mg/ATP mixture and terminated by adding 0.5% phosphoric acid after 40 min of incubation at room temperature. 10 μL of the reaction mixture was then added dropwise on a P30 filter pad and washed three times with 0.425% phosphoric acid and once with methanol within 4 minutes, then dried and analyzed using a scintillation counter. |
| Affinity data for this assay | |
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