| Assay Method Information | |
| | HTRF-Based Nucleotide Exchange Assay Detecting GTP Binding to K-Ras |
| Description: | A HTRF-based nucleotide exchange assay is used to characterize the pharmacological properties of the synthesized inhibitor compounds and the IC50 measurement is performed as described below. Kras G12D was mixed with a-GST Tb antibody (1.5× solution) and 10 uL was delivered to reaction wells. A mixture of Kras G12D/aGST-Tb Ab/compound mixture was incubated for 1 h at RT. A 3× solution of SOS1 (aa 564-1049) and fluorescently labelled GTP* was prepared in reaction buffer and 5 μL of SOS1/GTP* solution was added to reaction well to initiate the exchange reaction. An IC50 was determined using a sigmoidal dose response (variable slope) equation. |
| Affinity data for this assay | |
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