Treat MCF10A cells (in duplicate) with dilution series of LJH685 (Stock in DMSO): 100µM, 33µM, 10µM, 3µM, 1µM, 0.3µM, 0.1µM, 0µM (control) +/- 3ng/ml EGF for 30 min. Total of 32 samples. Extract and quantify activated Ras activity levels by ELISA.
Expected results: We expect to see an increase in Ras activity as we increase the concentrations of LJH685. We should see this increase in both the presence and absence of EGF but see a much stronger effect with EGF. The 30 min sample time should be enough time to reach signaling steady state, but not long enough to impact transcriptionally controlled feedback regulators. I suspect that 10µM is enough for maximum inhibition, but this might depend on cell type.
SEEK ID: https://emsl-seek.pnnl.gov/assays/21
Investigation: hidden item
Study: hidden item
Assay type: Inhibitor study
Technology type: ELISA
Created: 29th Oct 2021 at 00:51
Last updated: 8th Dec 2021 at 22:44