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Competitive Fluorescent Ligand Assay for Inositol 1,4,5-Trisphosphate

  • Akihiko TanimuraEmail author
  • Satoshi Shuto
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2091)

Abstract

We present a novel method, termed competitive fluorescent ligand assay for inositol 1,4,5-trisphosphate (CFLA-IP3), to measure inositol 1,4,5-trisphosphate (IP3). This method is based on fluorescence resonance energy transfer (FRET) between two fluorescent molecules, a fluorescent IP3-binding protein and its fluorescent ligand. Binding of these fluorescent molecules generates a FRET signal, and the IP3-dependent decrease in the FRET signal due to displacement of the fluorescent ligand is detected by fluorescence microscopy.

Key words

Inositol 1,4,5-trisphosphate IP3 receptor Ligand binding domain Fluorescent ligand Fluorescence resonance energy transfer Fluorescence microscopy 

Notes

Acknowledgments

This study was partly supported by the MEXT/Japan Society for the Promotion of Sciences (JSPS) KAKENHI under Grant No. JP16H01433 “Resonance Bio” and Grant No. 26670812 (to A.T.).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Division of Pharmacology, School of DentistryHealth Sciences University of HokkaidoHokkaidoJapan
  2. 2.Laboratory of Organic Chemistry for Drug Development, Faculty of Pharmaceutical SciencesHokkaido UniversityHokkaidoJapan

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