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Meten Eiwit Stabiliteit in Living zebravis embryo's met behulp van fluorescentie Decay Na Photoconversion (FDAP)
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Biologia do Desenvolvimento
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JoVE Journal Biologia do Desenvolvimento
Measuring Protein Stability in Living Zebrafish Embryos Using Fluorescence Decay After Photoconversion (FDAP)
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Meten Eiwit Stabiliteit in Living zebravis embryo's met behulp van fluorescentie Decay Na 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

Capítulos

  • 00:05Título
  • 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

Tadução automática

Tadução automática

Eiwitniveaus in cellen en weefsels vaak zijn gereguleerd door het evenwicht van eiwitproductie en klaring. Met behulp van fluorescentie Decay Na Photoconversion (FDAP), kan de klaringskinetiek van eiwitten experimenteel worden gemeten 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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