| Assay Method Information | |
| | Inhibition Test of Compounds on KRAS G12C::SOS1 Binding |
| Description: | Table 2: The compound to be tested was prepared into a 10 mM stock solution with DMSO, and serially diluted using 1× test buffer (modified Tris buffer). The resulting compound solution was transferred to a 384-well plate, and the final content of DMSO was 0.25%, and an additional DMSO well without compound was set up as a high signal control.1× test buffer (modified Tris buffer) was prepared. KRAS G12C (SignalChem, Cat. No. R06-32DH-BULK), SOS1 (Cytoskeleton, Inc., Cat. No. GE02-XL) and GTP solution (BellBrook, Cat. No. 3014-1K) were prepared respectively using the test buffer. 10 μL of KRAS G12C was transferred to the 384-well plate, and another 10 μL of the buffer was transferred to an empty well as a low signal control. 5 μL of SOS1 and 5 μL of GTP were transferred to the 384-well plate, respectively.GDP detection reagent was prepared using the test buffer. 10 μL of the detection reagent solution was transferred to the 384-well plate and incubated at room temperature for 2 h. After incubation, the plate was read on a multifunctional microplate reader (SpectraMax Paradigm) with an excitation wavelength of 580 nm and an emission wavelength of 620 nm. |
| Affinity data for this assay | |
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