| Assay Method Information | |
| | Determination of Allosteric Inhibition Activities of the Compounds of the Present Invention on SHP2 |
| Description: | he experimental process was briefly described as follows: Test compounds were first dissolved in DMSO to prepare storage solutions. The reaction was carried out in a 384-well Small Volume HiBase microplate (Greiner, 784075). Firstly, SHP2 (signalchem, P38-20G-10ug) and SHP-2 Activating Peptide (IRS1_pY1172(dPEG8)pY1222) BPS, 79319-1) were added to the wells till the final concentrations were 0.5 nM and 0.5 uM, respectively. Then, the compounds to be tested were added in a concentration range of 0.00004-10 uM and incubated at 25 C. for 60 minutes. Then DiFMUP (Thermo, D6567) was added in the reaction and incubated at 25 C. for 30 minutes. After incubation, readings were taken using a microplate reader (BMG) with excitation and emission wavelengths of 340 nm and 450 nm, respectively. Compared with the fluorescence intensity ratio of a control group (0.1% DMSO), the percentage inhibition rates of the compounds at each concentration were calculated, and the IC50 values of the compounds were obtained by performing nonlinear regression analysis with logarithmic value of the compound concentration inhibition rate by GraphPad Prism 5 software. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |