| Assay Method Information | |
| | RET Inhibition Experiment |
| Description: | The three drug-resistant mutant RET enzymes were each pre-incubated with different concentrations of test compounds (in the inhibition rate test of the compound: for RET-G810R and RET-G810C, the compound concentration was 300 and 1,000 nM; for RET-G810S, the compound concentration was 30 and 300 nM. In the IC50 test of the compound: for RET-G810R and RET-G810C, the compound concentration was 1-10,000 nM; for RET-G810S, the compound concentration was 0.1-10,000 nM, 9 concentrations in each case). After pre-incubation for 30 min at room temperature, the substrate and adenosine triphosphate (ATP) were added to start the reaction. For RET-G810R and RET-G810S, incubation was carried out at room temperature for 90 min; for RET-G810C, incubation was carried out at room temperature for 120 min. TK antibody-cryptate and streptavidin-XL665 were added, and the test was performed after incubation at room temperature for 60 min. |
| Affinity data for this assay | |
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