| Assay Method Information | |
| | Inhibitory Effects of Compounds on KRAS-G12C/SOS1 |
| Description: | Experimental reagents: KRASG12C/SOS binding kit (Cisbio, cat. 63ADK000CB16PEG); DMSO (Sigma, cat. D8418-1L); and 384-well white plate (PerkinElmer, cat. 6007290)Experimental Method1. Preparation of compounds: the test compounds were dissolved in 100% DMSO to obtain 10 mM stock solutions and the stock solutions were stored in a refrigerator in the dark.2. Kinase reaction process:Preparation of compounds: the test compounds had the concentration of 5,000 nM and diluted in a 384-well plate into a 100% DMSO solution at the 200-fold final concentration, and the compounds were diluted 3-fold with 10 concentrations. 50 nL of the compounds at the 200-fold final concentration were transferred to the plate of interest, the 384-well plate, using a dispenser Echo 550. 50 nL of 100% DMSO was added to each negative control well and positive control well.A Tag1-SOS1 solution at the 4-fold final concentration was prepared using a diluent buffer.2.5 μL of the Tag1-SOS1 solution at the 4-fold final concentration was added to the 384-well plate.A Tag2-KRAS-G12C solution at the 4-fold final concentration was prepared using the diluent buffer.2.5 μL of the Tag2-KRAS-G12C solution at the 4-fold final concentration was respectively added into compound wells and the positive control wells; and 2.5 μL of the diluent buffer was added to the negative control wells.The 384-well plate was centrifuged at 1,000 rpm for 30 seconds, shaken and mixed uniformly, and incubated at room temperature for 15 minutes.An Anti-Tag1-TB3+ solution at the 1-fold final concentration and an Anti-Tag2-XL665 solution at the 1-fold final concentration were prepared using a detection buffer, the two solutions were mixed uniformly to obtain a Mix solution, and 5 μL of the Mix solution was added into each well. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |