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Assessing Protein Interactions in Live-Cells with FRET-Sensitized Emission
JoVE Journal
Biologia
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JoVE Journal Biologia
Assessing Protein Interactions in Live-Cells with FRET-Sensitized Emission
DOI:

09:15 min

April 22, 2021

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Capítulos

  • 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

Tadução automática

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.

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