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Voltage-clamp Fluorometry in Xenopus Oocytes Using Fluorescent Unnatural Amino Acids
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Voltage-clamp Fluorometry in Xenopus Oocytes Using Fluorescent Unnatural Amino Acids
DOI:

09:24 min

May 27, 2017

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Chapters

  • 00:05Title
  • 01:03Oocyte Preparation and Injection
  • 04:16VCF Setup
  • 05:35VCF Recording
  • 07:44Results: Two-color VCF with Anap and TMR
  • 08:27Conclusion

Summary

Automatic Translation

This article describes an enhancement of conventional Voltage-Clamp Fluorometry (VCF) where Fluorescent Unnatural Amino Acids (fUAA) are used instead of maleimide dyes, to probe structural rearrangements in ion channels. The procedure includes Xenopus oocyte DNA injection, RNA/fUAA coinjection, and simultaneous current and fluorescence measurements.

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