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Single-Cell Quantification of Protein Degradation Rates by Time-Lapse Fluorescence Microscopy in Adherent Cell Culture
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Biologia dello sviluppo
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JoVE Journal Biologia dello sviluppo
Single-Cell Quantification of Protein Degradation Rates by Time-Lapse Fluorescence Microscopy in Adherent Cell Culture

Single-Cell Quantification of Protein Degradation Rates by Time-Lapse Fluorescence Microscopy in Adherent Cell Culture

DOI:
10.3791/56604-v

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12:06 min

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February 04, 2018

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,
1UPSUTER, Institute of Bioengineering (IBI), School of Life Sciences,École Polytechnique Fédérale de Lausanne (EPFL)

Capitoli

  • 00:00Titolo
  • 01:42Cell seeding and pulse labeling of the SNAP-tag
  • 04:15Time-lapse microscopy
  • 05:23Image processing and analysis
  • 07:55Risultati
  • 10:13Conclusions

Summary

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This protocol describes a method to determine protein half-lives in single living adherent cells, using pulse labeling and fluorescence time-lapse imaging of SNAP-tag fusion proteins.

Tags

Protein DegradationSingle-cellTime-lapse Fluorescence MicroscopyAdherent Cell CultureSNAP-tagProtein Half-lifeFluorescence ImagingProtein TurnoverHeterogeneityEndogenous Protein TaggingOverexpressionDNA Repair EnzymeBenzylguanineFluorescent DyeExponential DecayEmbryonic Stem CellsMonolayerCell-cell InteractionsN-cadherin

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