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Mätning Protein Stabilitet i Living Zebrafish embryon Använda Fluorescens Decay Efter Photoconversion (FDAP)
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Developmental Biology
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JoVE Journal Developmental Biology
Measuring Protein Stability in Living Zebrafish Embryos Using Fluorescence Decay After Photoconversion (FDAP)
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Mätning Protein Stabilitet i Living Zebrafish embryon Använda Fluorescens Decay Efter Photoconversion (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

Chapters

  • 00:05Title
  • 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

Automatic Translation

Automatic Translation

Protein nivåer i celler och vävnader är ofta hårt reglerad av balansen mellan produktionen och clearance-protein. Använda Fluorescens Decay Efter Photoconversion (FDAP), kan släppnings kinetiken av proteiner experimentellt mätas 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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