| Assay Method Information | |
| | Comparison of Selectivity Towards the JAK Family |
| Description: | 1. Experimental Method1.1. Pseudokinase (JH2) Experimental Operation was as Follows:1.1.1. Dissolve the compound with DMSO to a storage concentration of 10 mM.1.1.2. Prepare a compound with a concentration of 200 times the final concentration in a compound dilution plate by diluting it from the highest concentration point for a total of 4 concentration points using a 27 times dilution method, and transfer it to the Echo plate.1.1.3. Use an Echo instrument to pulse the compound from the Echo plate to a 384 experimental plate, thereby obtain the compound with 11 concentration points of the 3-fold dilution matrix.1.1.4. Add 5 ul 3×TYK2-JH2 or JAK1-JH2 pseudokinase to the 384-well experimental plate.1.1.5. Add 5 ul 3×Tb to the 384-well experimental plate.1.1.6. Add 5 ul 3× Tracer to the 384-well experimental plate.1.1.7. Centrifuge for 30 seconds and incubate at room temperature for 60 minutes.1.1.8. Read the signal value using Envision Microplate Reader (PerkinElmer). |
| Affinity data for this assay | |
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