| Assay Method Information | |
| | KRAS G12C-GDP Exchange Assay |
| Description: | 1. Serially 4×-diluted compounds (10 concentration points in total) were separately premixed with KRAS G12C-GDP (ICE, Kras 20191018) in a reaction buffer (25 mM Hepes PH7.4, 125 mM NaCl, 5 mM MgCl2, 0.01% Tween20, and 0.1% BSA) in the reaction wells for 1 h. 2. A mixture of SOS (Pharmaron, ZZY-20190823), cRAF (Pharmaron, ZZY-20190823), GTP (Sigma, A6885-100MG), MAb Anti 6HIS-d2/MAb Anti GST-Eu (Cisbio, 61HISDLB/61GSTKLB) was added for a catalyzed reaction for 2 h.3. The fluorescence signals of the emitted light at 615 nm and 665 nm under 320 nm excitation light were read by Biotek Microplate Reader (Synergy4).4. The IC50 (median inhibitory concentration) of the compound was obtained by the following non-linear fitting formula: Y=Bottom+(Top−Bottom)/(1+10{circumflex over ( )}((Log IC50−X)*Hill Slope)), and the data were analyzed by Graphpad 6-0 software. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |