MAE 6291 Lecture 06: Single-Molecule Fluorescence, TIRF, FRET

By jonathan silver

George Washington University, Washington, DC

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References

  1. A. Jain et al., Probing cellular protein complexes using single-molecule pull-down. Nature 474: 484-489 (2012)

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Researchers should cite this work as follows:

  • jonathan silver (2013), "MAE 6291 Lecture 06: Single-Molecule Fluorescence, TIRF, FRET," https://nanohub.org/resources/18850.

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George Washington University

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  1. nano/bio
MAE 6291 Lecture 06: Single-Molecule Fluorescence, TIRF, FRET
  • Single molecule pull-down 1. Single molecule pull-down 0
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  • What is fluorescence? 2. What is fluorescence? 249.98331664998332
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  • What is TIRF microscopy? 3. What is TIRF microscopy? 429.36269602936272
    00:00/00:00
  • What is photobleaching? bulk sample -> 4. What is photobleaching? bulk s… 792.85952619285956
    00:00/00:00
  • Model system to see captured protein 5. Model system to see captured p… 1106.6066066066067
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  • Results - Model system to see captured protein 6. Results - Model system to see … 1716.016016016016
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  • Monomer YFP dimeric YFP 7. Monomer YFP dimeric YFP 2058.5585585585586
    00:00/00:00
  • Results - Model system to see captured protein 8. Results - Model system to see … 2196.5965965965966
    00:00/00:00
  • Results - Model system to see captured protein 9. Results - Model system to see … 3098.1648314981649
    00:00/00:00
  • Can you give an estimate for the KD? 10. Can you give an estimate for t… 3109.542876209543
    00:00/00:00
  • Results - Model system to see captured protein 11. Results - Model system to see … 3826.6266266266266
    00:00/00:00
  • What is dynamic range of assay? 12. What is dynamic range of assay… 4032.4657991324661
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  • Use pull-down to study interaction between regulatory R and catalytic C proteins that form a complex 13. Use pull-down to study interac… 4144.7113780447116
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  • C captured via HA C captured via FLAG on R 14. C captured via HA C captured v… 4353.1865198531868
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  • acceptor 15. acceptor 4373.6069402736075
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  • What is fluorescence? 16. What is fluorescence? 4493.1931931931931
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  • acceptor 17. acceptor 4563.7971304637977
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  • X 18. X 5062.7961294627967
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  • Summary 19. Summary 5120.286953620287
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