| Assay Method Information | |
| | Tests of Compounds for Inhibiting HPK1 Kinase Activity In Vitro |
| Description: | The compound dilution was transferred into 384-well plate by using a pipette, afterwards, the 384-well plate was sealed and centrifuged at 1000 g for 1 min. 2-fold concentration of HPK1 solution was prepared in the kinase buffer, and then 2.5 μL of the prepared 2-fold concentration of HPK1 solution was added into the 384 wells, centrifuged at 1000 g for 30 s, and incubated for 10 min at room temperature. A mixture of 2-fold concentration of MBP and ATP was prepare in the kinase buffer, and 2.5 sL of prepared mixture of 2-fold concentration of MBP and ATP was added into the above reaction system for the reaction, centrifuged at 1000 g for 30 s, and then incubated at room temperature for 1 h, 5 μL of ADP-Glo reagent was added into the reaction system, and incubated at room temperature for 40 min. 10 μL of kinase detection reagent was added and incubated at room temperature for 40 min. Then the luminescence signals were read on an Envision 2104 plate reader. |
| Affinity data for this assay | |
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