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Monitoring Kinase and Phosphatase Activities Through the Cell Cycle by Ratiometric FRET
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Biologia
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JoVE Journal Biologia
Monitoring Kinase and Phosphatase Activities Through the Cell Cycle by Ratiometric FRET

Monitoring Kinase and Phosphatase Activities Through the Cell Cycle by Ratiometric FRET

DOI:
10.3791/3410-v

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13:38 min

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January 27, 2012

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, , ,
1Department of Cell and Molecular Biology,Karolinska Institutet

Capitoli

  • 00:05Titolo
  • 02:19Introducing the Probe to Cells
  • 03:27Monitoring Ratiometric FRET
  • 06:12Verifying and Optimizing Conditions
  • 06:43Analyzing FRET
  • 11:24Representative Imaging Results
  • 12:52Conclusion

Summary

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FRET-based reporters are increasingly used to monitor kinase and phosphatase activities in live cells. Here we describe a method on how to use FRET-based reporters to assess cell cycle-dependent changes in target phosphorylation.

Tags

KinasePhosphataseCell CycleRatiometric FRETFörster Resonance Energy TransferFRET-based ReportersCFPYFPPhosphorylationConformation ChangeDistanceOrientationFRET EfficiencyProbesPKAPKBPKCPKDERKJNKCdk1Aurora BPlk1Stress Signaling PathwaysProgression Through The Cell CycleUnperturbed GrowthRecovering From StressTime-lapse ImagingSignal To Noise Ratio

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