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In Vivo Quantification of Protein Turnover in Aging C. Elegans using Photoconvertible Dendra2
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In Vivo Quantification of Protein Turnover in Aging C. Elegans using Photoconvertible Dendra2

In Vivo Quantification of Protein Turnover in Aging C. Elegans using Photoconvertible Dendra2

DOI:
10.3791/61196-v

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09:45 min

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June 13, 2020

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,
1Leibniz Research Institute for Molecular Pharmacology im Forschungsverbund, 2NeuroCure Cluster of Excellence,Charité – Universitätsmedizin Berlin, 3Department of Biology and Chemistry,University of Bremen

Chapters

  • 00:04Introduction
  • 00:50Microscope Slide Preparation
  • 01:38Confocal Microscope Setup
  • 03:17C. elegans Mounting
  • 04:06Green Dendra2 Conversion: Data Acquisition at Time Zero
  • 05:35Red Dendra2 Conversion: Data Acquisition at Second Time Point
  • 06:13Converted Dendra2 Image Analysis
  • 07:17Results: Representative Huntingtin (HTT)-D2 Conversion and Degradation
  • 09:08Conclusion

Summary

자동 번역

자동 번역

Presented here is a protocol to monitor degradation of the protein huntingtin fused to the photoconvertible fluorophore Dendra2.

Tags

Keywords: Protein TurnoverDendra2C. ElegansPhotoconversionIn VivoQuantificationAgingProteostasisTraffickingMicroscopyAgarose PadConfocalLaserAcquisition SettingsConversionBleaching

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