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Exploring Muscarinic Acetylcholine Receptor Binding Kinetics with Fluorescence Anisotropy

  • Tõnis Laasfeld,
  • Maris-Johanna Tahk,
  • Anni Allikalt,
  • Jane Torp,
  • Lukas Grätz,
  • Sergei Kopanchuk,
  • Ago Rinken

摘要

Many different assays are available for measuring ligand binding to muscarinic acetylcholine receptors. Each has advantages and throwbacks, making the optimal choice for a specific research question nontrivial. This chapter gives a detailed description and protocol of a fluorescence anisotropy assay, which enables real-time monitoring of fluorescently labelled and non-labelled ligand binding to untagged receptors. Depending on the goal of the respective study, we further provide instructions for data analysis on three different complexity levels along with example data and a repository of software and data files for convenient application of the explained data analysis techniques