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Assessing Protein Interactions in Live-Cells with FRET-Sensitized Emission
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Assessing Protein Interactions in Live-Cells with FRET-Sensitized Emission

Assessing Protein Interactions in Live-Cells with FRET-Sensitized Emission

DOI:
10.3791/62241-v

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09:15 min

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April 22, 2021

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, , , , ,
1Department of Biophysics and Cell Biology, Faculty of Medicine,University of Debrecen, 2Gynecologic Oncology Division,Stanford University School of Medicine

Chapters

  • 00:05Introduction
  • 01:12Plasmid Construction
  • 01:52Cell Culture and Transfection
  • 02:41FRET Imaging
  • 05:42Image Analysis for Detecting Absolute FRET Efficiencies Using Donor Quenching and Sensitized Emission
  • 07:01Results: Quantification of FRET Images
  • 08:28Conclusion

Summary

Automatic Translation

Automatic Translation

Förster Resonance Energy Transfer (FRET) between two fluorophore molecules can be used for studying protein interactions in the living cell. Here, a protocol is provided as to how to measure FRET in live cells by detecting sensitized emission of the acceptor and quenching of the donor molecule using confocal laser scanning microscopy.

Tags

Keywords: FRETFRET-sensitized EmissionProtein InteractionsLive-cell ImagingDonor QuenchingAcceptor Sensitized EmissionCrosstalkFluorescent ProteinsDetection EfficiencyDonor-acceptor Fusion ProteinEGFP-mCherryCell TransfectionConfocal MicroscopyEnvironmental Chamber

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