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Novel Tools for Use in Bioluminescence Resonance Energy Transfer (BRET) Assays

  • Mélanie Robitaille
  • Isabelle Héroux
  • Alessandra Baragli
  • Terence E. Hébert
Protocol
Part of the Methods in Molecular Biology™ book series (MIMB, volume 574)

Abstract

Recent advances in imaging assays based on bioluminescence resonance energy transfer (BRET) have made it possible to study protein/protein interactions in living cells under physiological conditions. Here we describe protocols for these assays including relevant positive and negative controls, and we also show how they can be combined with protein complementation assays such as bimolecular fluorescence complementation (BiFC) to study three- and four-partner interactions. We also describe a BRET assay that uses SNAP-tagged proteins as a fluorescence acceptor molecule for the bioluminescent donor.

Key words

BRET GPCRs SNAP tagging bioluminescence biomolecular fluorescence complementation 

Notes

Acknowledgments

This work was supported by grants from the Canadian Institutes of Health Research (MOP-99567) and Heart and Stroke Foundation of Quebec to T.E.H. T.E.H. is a Chercheur National of the Fonds de Recherche en Santé du Québec (FRSQ). M.R. holds a doctoral scholarship from the FRSQ. We thank Vic Rebois (NIH) for helpful discussions and we also thank Covalys for providing constructs and reagents for the SNAP tag.

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Mélanie Robitaille
    • 1
  • Isabelle Héroux
    • 1
  • Alessandra Baragli
    • 2
  • Terence E. Hébert
    • 2
  1. 1.Département de Biochimie, Department of Pharmacology and TherapeuticsUniversité de Montréal, McGill UniversityMontréalCanada
  2. 2.Department of Pharmacology and TherapeuticsMcGill UniversityMontréalCanada

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