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

•

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

Bölümler

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

Özet

Otomatik Çeviri

Otomatik Çeviri

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.

Etiketler

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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