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Visualiseren Clathrine-gemedieerde endocytose van G-proteïne gekoppelde receptoren bij Single-event resolutie via TIRF Microscopie
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Biologia
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JoVE Journal Biologia
Visualizing Clathrin-mediated Endocytosis of G Protein-coupled Receptors at Single-event Resolution via TIRF Microscopy
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Visualiseren Clathrine-gemedieerde endocytose van G-proteïne gekoppelde receptoren bij Single-event resolutie via TIRF Microscopie

DOI:
10.3791/51805-v

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12:40 min

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October 20, 2014

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, ,
1Department of Biological Sciences,Carnegie Mellon University

Capítulos

  • 00:05Título
  • 01:21Expression of Flourescently Tagged CME Components in Cultured Cells
  • 02:42Imaging Clathrin-coated Pit (CCP) and Cargo Dynamics Using TIRFM
  • 05:22Analysis of Endocytic Dynamics by Manual Verification
  • 06:32Analysis of Endocytic Dynamics by Objective Recognition
  • 10:40Results: Anlysis of the Endocytic Dynamics of the μ‐opioid‐receptor (MOR), a GPCR, and its Endocytic Adaptor Protein β‐arrestin
  • 12:18Conclusion

Summary

Tadução automática

Tadução automática

Clathrin-mediated endocytosis, a rapid and highly dynamic process internalizes many proteins, including signaling receptors. The protocol described here directly visualizes the kinetics of individual endocytic events. This is essential for understanding how core members of the endocytic machinery coordinate with each other, and how protein cargo influence this process.

Tags

Keywords: Clathrin-mediated EndocytosisG Protein-coupled ReceptorsTIRF MicroscopySingle-event ResolutionEndocytic MachineryClathrin-coated PitsCargo CompositionFluorescence ProbesReal-time VisualizationCell Biology

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