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A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts
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Chapters
- 00:04Title
- 00:49Sample Preparation: Cell-Cell Mixing Assay
- 02:37Scanning Fluorescence Cross-Correlation Spectroscopy (sFCCS): Acquisition
- 04:07Scanning Fluorescence Cross-Correlation Spectroscopy (sFCCS): Data Analysis
- 06:03Results: Investigation of Protein-Protein Interactions at Cell-Cell Contacts by sFCCS
- 08:09Conclusion
This protocol describes a fluorescence fluctuation spectroscopy-based approach to investigate interactions among proteins mediating cell-cell interactions, i.e. proteins localized in cell junctions, directly in living cells. We provide detailed guidelines on instrument calibration, data acquisition and analysis, including corrections to possible artefact sources.
Tags
Keywords:
Fluorescence Fluctuation SpectroscopyProtein-protein InteractionsCell-cell ContactsCell AdhesionCell RecognitionLiving CellsMono-membrane VesiclesModel OrganismsCell TransfectionGreen Fluorescent ProteinYellow Fluorescent ProteinRed Fluorescent ProteinEDTACell DetachmentCell ResuspensionLaser Scanning Confocal MicroscopeSpectral CrosstalkMEGFPMEYFPMCherryMCardinal
