| Assay Method Information | |
| | PI3Kα[WT]/[E542K]/[E545K]/[H1047R]/PI3Kβ/γ/δ Enzyme Activity Assay (ADP Glo) |
| Description: | PI3Kα[WT], PI3Kα[E542K], PI3Kα[E545K], PI3Kα[H1047R], PI3Kβ, PI3Kγ and PI3Kδ kinase reaction was performed in 384-well plate (PerkinElmer, #6008280), with a reaction system of 4 μL. PI3Kα[WT] enzyme solution (Promega, #V1721) or PI3Kα[E542K] enzyme solution (CarnaBio, #11-413-20N) or PI3Kα[E545K] enzyme solution (CarnaBio, #11-414-20N) or PI3Kα[H1047R] enzyme solution (Promega, #V1741) or PI3Kβ enzyme solution (Promega, #V1751) or PI3Kγ enzyme solution (ThermoFisher, #PV4786) or PI3Kδ enzyme solution (ThermoFisher, #PV6451) were added to 384-well plate, and then gradiently diluted compounds were added (starting at a final concentration of 10 μM, 3-fold dilution, 10 doses), followed by adding PIP2: 3PS (Promega, #V1701) and ATP solution (Promega, #V915B) to the plate. After incubation at 25° C. for 1 h, ADP-Glo reagent buffer (Promega, #V9102) was added to each well. The plate was sealed and incubated at 25° C. for 40 min, and then 10 μL of ADP-Glo detection buffer (Promega, #V9102) was added to each well. After incubating at 25° C. for 40 min, the chemiluminescence value was read using an instrument. |
| Affinity data for this assay | |
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