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Visualizing the Conformational Dynamics of Membrane Receptors Using Single-Molecule FRET
JoVE Journal
生化学
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JoVE Journal 生化学
Visualizing the Conformational Dynamics of Membrane Receptors Using Single-Molecule FRET

Visualizing the Conformational Dynamics of Membrane Receptors Using Single-Molecule FRET

DOI:

10:59 min

August 17, 2022

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  • 00:04Introduction
  • 01:04Slide and Coverslip Cleaning
  • 03:04mGluR2 Expression with Incorporated Unnatural Amino Acid, Fluorescent Labeling, and Extraction
  • 05:13Single-Molecule Flow Chamber Assembly and Functionalization
  • 06:23Microscope Setup and smFRET Data Acquisition
  • 07:47Data Analysis
  • 08:39Results: Analyzing Conformational Dynamics of Membrane Receptors Proteins
  • 10:11Conclusion

概要

自動翻訳

This study presents a detailed procedure to perform single-molecule fluorescence resonance energy transfer (smFRET) experiments on G protein-coupled receptors (GPCRs) using site-specific labeling via unnatural amino acid (UAA) incorporation. The protocol provides a step-by-step guide for smFRET sample preparation, experiments, and data analysis.

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