| Assay Method Information | |
| | Inhibitory Activities of the Compounds on JAK1, JAK2, JAK3 and Tyk2 (IC50) |
| Description: | A compound (powder) was dissolved in 100% DMSO to prepare a 10 mM storage solution. The compound had an initial test concentration of 10,000 nM or 1000 nM, was 3-fold or 4-fold serially diluted to obtain 10 concentrations. Detection in duplicate. Baricitinib (Selleckchem, Cat. S2851) was used as positive control. The serially diluted compound was mixed with JAK1/JAK2/JAK3/Tyk2 kinase (Cama, Cat. 08-144/08-045/08-046/08-147) with final concentration of 5 nM/0.125 nM/0.5 nM/2.5 nM in a Optiplate-384F plate and incubated at room temperature for 10 minutes. After that, ATP with final concentration of 1 mM and 3 μM Kinase Substrate 30 (GL Biochem, Cat. 117885) were added and mixed well. The reaction was carried out at room temperature for 20 min. Stop test solution was added to stop the reaction and the conversion rate was read by Caliper EZ Reader II. |
| Affinity data for this assay | |
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