| Assay Method Information | |
| | Inhibition of KIF18A Enzyme Activity |
| Description: | The ADP-Glo method was used to test the enzymatic activity of KIF18A. The compound was diluted 3-fold with DMSO at a starting concentration of 3000 nM, for a total of ten concentrations, and tested in duplicate. Echo was used to transfer 25 nL of the diluted compound to a 384-well plate, and 2.5 μL of enzyme working solution was added. Plate was centrifuged at 1000 rpm for 1 min, and incubated at 25° C. for 10 min. 2.5 μL of ATP and substrate solution was added to start the reaction, and plate was incubated at 25° C. for 60 min. 4 μL of ADP-Glo solution was added to the 384-well plate, and plate was incubated at 25° C. for 40 min. 8 μL of detection solution was added and incubated at 25° C. for 40 min, and BMG was used to read the fluorescence value. Inhibition rate was calculate according to the following formula: Inhibition rate (%)=(DMSO well−compound well)/(DMSO well−blank control well)×100%. The IC50 value of each compound was analyzed using the nonlinear regression method of XLFit 5.5.0 software. The formula is: Y=Bottom+(Top−Bottom)/(1+10{circumflex over ( )}((Log IC50−X)*HillSlope)). |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |