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Assays for the Degradation of Misfolded Proteins in Cells
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JoVE Journal Biology
Assays for the Degradation of Misfolded Proteins in Cells

Assays for the Degradation of Misfolded Proteins in Cells

DOI:
10.3791/54266-v

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10:56 min

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August 28, 2016

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, ,
1Department of Cancer Biology,University of Pennsylvania Perelman School of Medicine, 2Department of Systems Pharmacology and Translational Therapeutics,University of Pennsylvania Perelman School of Medicine

Chapters

  • 00:05Title
  • 01:04Degradation Assay of Atxn1 82Q GFP
  • 05:06Degradation Assay of NLS-luciferase-GFP Using SDS-PAGE and Western Blot
  • 06:42Real-time Degradation Assay of NLS-luciferase-GFP Using Fluorescence Microplate Reader
  • 07:38Results: Degradation of Misfolded Proteins
  • 09:15Conclusion

Summary

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This report describes protocols for measuring degradation rates of misfolded proteins by either western blot or fluorescence-based assays. The methods can be applied to analysis of other misfolded proteins and for high throughput screening.

Tags

Misfolded ProteinsProtein DegradationProtein Quality ControlNeurodegenerative DiseasesAtxn1 82QNuclear LuciferaseCell FractionationCycloheximideProteasome InhibitorMG132Cell LysisCentrifugation

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