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Måling Protein Stabilitet i Living zebrafisk embryoner anvendelse af fluorescens Decay Efter fotoomdannelse (FDAP)
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Journal JoVE Biologie du développement
Measuring Protein Stability in Living Zebrafish Embryos Using Fluorescence Decay After Photoconversion (FDAP)

Måling Protein Stabilitet i Living zebrafisk embryoner anvendelse af fluorescens Decay Efter fotoomdannelse (FDAP)

DOI:
10.3791/52266-v

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09:45 min

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January 28, 2015

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, , ,
1Department of Molecular and Cellular Biology,Harvard University, 2Systems Biology of Development Group,Friedrich Miescher Laboratory of the Max Planck Society

Chapitres

  • 00:05Titre
  • 01:27Generating a Photoconvertible Fusion Construct and Injecting Dechorionated Zebrafish Embryos
  • 03:22Mounting Zebrafish Embryos for Photoconversion and Confocal Imaging
  • 04:37Photoconverting and Measuring the Decrease of the Photoconverted Signal
  • 06:47Analyzing the Data Using PyFDAP
  • 08:18Results: Half-life of Squint-Dendra2 as Determined by Fluorescence Decay After Photoconversion during Embryogenesis
  • 09:19Conclusion

Summary

Traduction automatique

Traduction automatique

Protein niveauer i celler og væv er ofte stramt reguleret af balance proteinproduktion og clearance. Anvendelse af fluorescens Decay Efter fotoomdannelse (FDAP) kan clearance kinetik proteiner eksperimentelt målt in vivo.

Tags

Protein StabilityFluorescence Decay After PhotoconversionFDAPProtein Clearance KineticsPhotoconvertible Fluorescent ProteinZebrafish EmbryosCompartmental AnalysisIntra- And Extracellular Half-livesSecreted Signaling ProteinsIn Vivo Protein Dynamics

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