| Assay Method Information | |
| | JAK1 Kinase Inhibitory Activity Determination |
| Description: | 1) 4 μL of the 2.5× compound to be tested was added to an HTRF 96 well low volume plate, and then 2 μL of 5× substrate was added from one side of the wells, 2 μL of 5×JAK1 was added from the other side of the wells; 2 μL of 1× kinase buffer was added in the blank control group; 2) the plate was sealed with a plate sealing film and placed in a centrifuge for centrifuging at 1000 rpm for 2 minutes; 3) 2 μL of 5×ATP was added to each well, and the plate was sealed with a plate sealing film, centrifuged at 1000 rpm for 1 min, and then put into an incubator at 30° C. for incubating 4 h; 4) at the end of incubation, 1 μM Streptavidin-XL665 and 1×TK-Antibody described above were mixed at a ratio of 1:1, then added to each well with a volume of 10 μL; the plate was centrifuged at 1000 rpm for 1 minute; 5) the plate was then put back into the incubator for incubating another 1 h; at the end of incubation, HTRF665/620 signals were read on a multifunctional microplate reader. |
| Affinity data for this assay | |
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