| Assay Method Information | |
| | Enzymologic Experiment of FGFR4 |
| Description: | In this experiment, the inhibitory effect of the compounds on FGFR4 kinase activity was tested by a fluorescence resonance energy transfer (TR-FRET) method, and the half maximal inhibitory concentration (IC50) of the compounds on the FGFR4 kinase activity was determined. 1) 1 5 μL of FGFR4 enzyme solution were added to a 384-well plate, and the final concentration of the enzyme was 0.1 5 nM. 2) 1 5 μL of diluted solution in gradient of the compound were added.3) 1 5 μL of a substrate mixture containing substrate polypeptide with a final concentration of 5 50 nM and ATP with a final concentration of 10 200 μM were added.4) The mixture was incubated for 0.5-3 hours at room temperature.5) 10 μL of EDTA and a test solution comprising labeled antibody were added, and the plate was incubated for 1 hour at room temperature.6) The fluorescence signal values of each plate were determined by a microplate reader at 665 nm.7) The inhibition rates were calculated according to the fluorescence signal values. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |