Journal
/
生物化学
/
Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells
JoVE 杂志
生物化学
Author Produced
This content is Free Access.
JoVE 杂志 生物化学
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

•

14:12 min

•

December 11, 2021

•

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

Chapters

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

Summary

自动翻译

自动翻译

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.

Tags

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

Article

Embed

添加到播放列表

Usage Statistics

Related Videos

Method for Identifying Small Molecule Inhibitors of the Protein-protein Interaction Between HCN1 and TRIP8b

Fluorescence Anisotropy as a Tool to Study Protein-protein Interactions

Using Scaffold Liposomes to Reconstitute Lipid-proximal Protein-protein Interactions In Vitro

Visualization of Protein-protein Interaction in Nuclear and Cytoplasmic Fractions by Co-immunoprecipitation and In Situ Proximity Ligation Assay

Biotinylated Cell-penetrating Peptides to Study Intracellular Protein-protein Interactions

Dissipative Microgravimetry to Study the Binding Dynamics of the Phospholipid Binding Protein Annexin A2 to Solid-supported Lipid Bilayers Using a Quartz Resonator

A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts

Using In Vitro Fluorescence Resonance Energy Transfer to Study the Dynamics Of Protein Complexes at a Millisecond Time Scale

Application of Biolayer Interferometry (BLI) for Studying Protein-Protein Interactions in Transcription

Quantification of Protein Interaction Network Dynamics using Multiplexed Co-Immunoprecipitation

Read Article