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Measuring Protein Stability in Living Zebrafish Embryos Using Fluorescence Decay After Photoconversion (FDAP)
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JoVE Journal Gelişim Biyolojisi
Measuring Protein Stability in Living Zebrafish Embryos Using Fluorescence Decay After Photoconversion (FDAP)

Measuring Protein Stability in Living Zebrafish Embryos Using Fluorescence Decay After 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

Bölümler

  • 00:05Başlık
  • 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

Özet

Otomatik Çeviri

Otomatik Çeviri

Protein levels in cells and tissues are often tightly regulated by the balance of protein production and clearance. Using Fluorescence Decay After Photoconversion (FDAP), the clearance kinetics of proteins can be experimentally measured in vivo.

Etiketler

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

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