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Ion Channels pp 217–231Cite as

The Use of Dansyl-Calmodulin to Study Interactions with Channels and Other Proteins

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 998))

Abstract

Steady-state fluorescence spectroscopy is a biophysical technique widely employed to characterize ­interactions between proteins in vitro. Only a few proteins naturally fluoresce in cells, but by covalently attaching fluorophores virtually all proteins can be monitored. One of the first extrinsic fluorescent probes to be developed, and that is still in use, is dansyl chloride. We have used this method to monitor the interaction of a variety of proteins, including ion channels, with the Ca2+-dependent regulatory protein calmodulin. Here we describe the preparation and use of dansyl-calmodulin (D-CaM).

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Acknowledgments

This work was supported by grants from the Spanish Ministry of Education (BFU2009-07581), the Spanish Ion Channel Initiative Consolider project (CSD2008-00005), and the Basque Government (SAIOTEK SA-2006/00023). A. Alaimo was partially funded by Fundación Biofísica Bizkaia. Proteomics Core Facility-SGIKER is a member of ProteoRed-ISCIII. Technical and human support provided by SGIKER (UPV/EHU, MICINN, GV/EJ, ERDF, and ESF) is gratefully acknowledged.

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Authors and Affiliations

  1. Unidad de Biofísica (CSIC-UPV/EHU), Universidad del País Vasco, Leioa, Spain

    Alessandro Alaimo, Covadonga Malo & Alvaro Villarroel

  2. Faculty of Medicine and Odontology, Department of Pharmacology, Universidad del País Vasco, Leioa, Spain

    Pilar Areso

  3. Proteomics Core Facility-SGIKER, Universidad del País Vasco, Leioa, Spain

    Kerman Aloria

  4. Proteomics Unit, CIC bioGUNE CIBERehd, Derio, Spain

    Oscar Millet

Authors
  1. Alessandro Alaimo
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  2. Covadonga Malo
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  3. Pilar Areso
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  4. Kerman Aloria
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  5. Oscar Millet
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  6. Alvaro Villarroel
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Editor information

Editors and Affiliations

  1. School of Biomedical Sciences, University of Leeds, Garstang Bld. 6.31d, Leeds, LS2 9JT, United Kingdom

    Nikita Gamper

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Alaimo, A., Malo, C., Areso, P., Aloria, K., Millet, O., Villarroel, A. (2013). The Use of Dansyl-Calmodulin to Study Interactions with Channels and Other Proteins. In: Gamper, N. (eds) Ion Channels. Methods in Molecular Biology, vol 998. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-351-0_17

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  • DOI: https://doi.org/10.1007/978-1-62703-351-0_17

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-350-3

  • Online ISBN: 978-1-62703-351-0

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