| Assay Method Information | |
| | KRAS G12D&SOS1 Binding Assay |
| Description: | The positive inhibitor and test compounds (10 mM storage solution) were diluted by a 3-fold gradient with 100% DMSO. Transfer 2 μL diluted compound to an 18 μL diluent and mix thoroughly. Take 2 μL of the diluted compound into a 384-well plate and repeated twice for each sample. Centrifuge 384 well plates at 1000 rpm and add 4 μL of KRAS G12D>P mixture into each well, followed by 4 μL of SOS1, and incubate for 15 minutes. Then 10 μL of Tag-Tb and Tag-XL665 were added and incubated at 4° C. for 3 hours. The HTRF (665 nM and 615 nM) signals were read using BMG. With the log value of concentration as the X-axis and 665 nM/615 nM as the Y-axis, Y=Bottom+(Top-Bottom)/(1+10∧((LogIC50-X)*HillSlope)) was used to fit the dose-effect curve. The IC50 value of each compound on enzyme activity was obtained. |
| Affinity data for this assay | |
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