| Assay Method Information | |
| | Protein-Protein Interaction (PPI) Assay |
| Description: | The assay was carried according to the manufacturer's instructions. Compounds at an initial concentration of 10 mM in DMSO were first serially diluted on a 384-well plate using assay buffer at a ratio of 1:3 for a total of 9 concentrations with each of a volume of 20 μL. Blank buffer was included as the negative control. The final compound solution was in the buffer solution with 1% DMSO. Then, in each well of a 384-well plate, a solution of 2 μL compound solution, 4 μL tagged SOS1 and KRAS (G12C) protein stocks, respectively, and 10 μM GTP was incubated for 15 min. 10 μL mixture of anti-Tag1X665 antibody and anti-Tag1-Terbium antibody was introduced to each individual well. After 2 hours incubation at room temperature, fluorescence readout was measured with a Pelkin Elmer multiple plate reader VICTOR Nivo. |
| Affinity data for this assay | |
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