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Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells
JoVE Journal
Bioquímica
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JoVE Journal Bioquímica
Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells

Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells

DOI:
10.3791/62954-v

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14:12 min

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December 11, 2021

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, , , , , ,
1Rudolf Virchow Center for Integrative and Translation Bioimaging,Julius-Maximilian University Wuerzburg, 2Max Delbrück Center for Molecular Medicine, Berlin

Capítulos

  • 00:05Introduction
  • 01:09Sample Preparation
  • 04:34Calibration Measurements
  • 05:58Live Cell Measurements
  • 06:52Data Export
  • 07:23Calibration Measurements
  • 09:20Live Cell Experiments
  • 11:20Resultados
  • 13:17Conclusion

Summary

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We present an experimental protocol and data analysis workflow to perform live cell dual-color fluorescence cross correlation spectroscopy (FCCS) combined with Förster Resonance Energy transfer (FRET) to study membrane receptor dynamics in live cells using modern fluorescence labeling techniques.

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Keywords: Fluorescence Correlation SpectroscopyCross-correlation SpectroscopyFRETG Protein-coupled ReceptorsBeta-2 Adrenergic ReceptorsSphingolipid ReceptorsProtein-protein InteractionProtein DynamicsLive CellsCell CultureTransfectionSNAP-tagFluorescence AnisotropyTranslational DiffusionRotational Diffusion

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