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A Semi-Quantitative Drug Affinity Responsive Target Stability (DARTS) assay for studying Rapamycin/mTOR interaction
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Chapters
- 00:04Title
- 00:47Collect and Lyse Cells
- 01:59Incubate Protein Lysates with the Small Molecule and Perform Proteolysis
- 03:28Western Blot Analysis
- 03:59Results: Rapamycin Inhibition
- 05:03Conclusion
In this study, we enhanced the data analysis capabilities of the DARTS experiment by monitoring the changes in protein stability and estimating the affinity of protein-ligand interactions. The interactions can be plotted into two curves: a proteolytic curve and a dose-dependence curve. We have used mTOR-rapamycin interaction as an exemplary case.
