Titrate the impact of LJH685 on Ras activation

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

Experimental assay

Steven Wiley

Projects: Reverse Sensitivity Analysis for Identifying Predictive Proteomics Signa...

Investigation: hidden item

Study: hidden item

Assay position:

Assay type: Inhibitor study

Technology type: ELISA

Organisms: Homo sapiens : MCF 10A (wild-type / Epithelial)

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Created: 29th Oct 2021 at 00:51

Last updated: 8th Dec 2021 at 22:44

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