| Assay Method Information | |
| | Ret wt and RET V804M kinase assay |
| Description: | In 5x kinase buffer solution A, Ret wt or RET V804M kinase was mixed with pre-formulated and diluted compounds of different concentrations for 10 minutes in duplicate for each concentration. The corresponding substrate and ATP were added thereto, and reacted at room temperature for 20 minutes (both a negative and a positive control were set: the negative control is blank, the positive control is CEP-32496). Detection reagent (reagent in HTRF KinEASE-TK kit) was added after the reaction is complete, after 30 minutes of incubation at room temperature, Envision ELISA reader was used for detection of enzyme activities in the presence of compounds of the present disclosure with different concentrations, and the inhibitory activities of different concentrations of compounds on enzyme activity were calculated, Graphpad 5.0 software was used to fit the inhibitory activities on enzyme activity against different concentrations of compounds, and IC50 values were calculated. |
| Affinity data for this assay | |
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