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Протокол по использованию Ферстер Resonance Energy Transfer (FRET) -Force биосенсоров для измерения механических сил по всему комплексу ядерного ЛИНКА
JoVE Journal
Biotechnik
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JoVE Journal Biotechnik
A Protocol for Using Förster Resonance Energy Transfer (FRET)-force Biosensors to Measure Mechanical Forces across the Nuclear LINC Complex
DOI:

09:43 min

April 11, 2017

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Kapitel

  • 00:05Titel
  • 00:53Transfect Cells with Nesprin-2G and Other Plasmid DNA
  • 04:09Verify Transfection Efficiency
  • 05:12Capture Spectral Fingerprints of mTFP1 and Venus Fluorophores for Spectral Unmixing
  • 06:53Capture Unmixed Images
  • 07:52Results: Setting Imaging Parameters and Capturing Spectrally Resolved FRET Images
  • 08:51Conclusion

Summary

Automatische Übersetzung

A number of FRET-based force biosensors have recently been developed, enabling the protein-specific resolution of intracellular force. In this protocol, we demonstrate how one of these sensors, designed for the linker of the nucleoskeleton-cytoskeleton (LINC) complex protein Nesprin-2G can be used to measure actomyosin forces on the nuclear LINC complex.

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