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Assay Development for High Content Quantification of Sod1 Mutant Protein Aggregate Formation in Living Cells
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Capítulos
- 00:05Título
- 00:58Lentivirus Production
- 02:50Lentiviral Transduction and Stable Line Generation
- 04:53Dose Response Effect of Proteosome Inhibitors on Protein Aggregation and Image Analysis
- 07:06Time lapse Imaging of YFP Expression in Living Cells
- 08:09Results: Quantification of SOD1 Mutant Protein Aggregate Formation
- 10:04Conclusion
We describe a method to quantify the aggregation of misfolded proteins. Our protocol details lentiviral induced stable cell line generation, automated confocal imaging, and image analysis of protein aggregates. As an illustrative application, we studied the effect of small molecules in promoting SOD1 aggregation in a time- and dose-dependent manner.
