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Assay Development for High Content Quantification of Sod1 Mutant Protein Aggregate Formation in Living Cells
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JoVE Journal Neuroscience
Assay Development for High Content Quantification of Sod1 Mutant Protein Aggregate Formation in Living Cells

Assay Development for High Content Quantification of Sod1 Mutant Protein Aggregate Formation in Living Cells

DOI:
10.3791/56425-v

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11:12 min

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October 04, 2017

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, , ,
1Automation & Logistics Management, Screening Sciences & Novel Assay Technologies,Institut Pasteur Korea, 2School of Pharmacy,Sungkyunkwan University, 3Technology Development Platform,Institut Pasteur Korea

Chapters

  • 00:05Title
  • 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

Summary

Automatic Translation

Automatic Translation

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.

Tags

Keywords: Assay DevelopmentHigh Content QuantificationSod1 Mutant ProteinProtein Aggregate FormationLiving CellsNeurodegenerative DisordersALSLentivirus ProductionHEK-293 T-cellsTransfectionCell Transduction

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