| Assay Method Information | |
| | Inhibition Test of Compounds on KRAS G12C::SOS1 Binding |
| Description: | Table 1: The compound to be tested was prepared into a 10 mM stock solution with DMSO, and the compound was serially diluted using 1× test buffer. 0.1 μL of the compound solution with different concentrations was transferred to a 384-well plate. 5 μL of GST-KRAS G12C was added to the 384-well plate, and centrifuged at 1000 rpm for 1 min. 5 μL of His-SOS1 was added to the 384-well plate, centrifuged at 1000 rpm for 1 min, and incubated at room temperature for 15 min.After incubation, 10 μL of a mixed solution of anti-6his-Tb monoclonal antibody (Cisbio, Cat. No. 61HI2TLA) and anti-GST-XL665 monoclonal antibody (Cisbio, Cat. No. 61GSTXLA) was added to the test well, centrifuged at 1000 rpm for 1 min, and incubated at room temperature for 1 h.After incubation, the fluorescence signal ratios at 665 nm and 615 nm were read on a multifunctional microplate reader (Perkin Elmer, Envision 2104), and the IC50 values were calculated using Graphpad 5 software. |
| Affinity data for this assay | |
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