Summary
Fluorescent dyes sensitive to changes in intracellular calcium have become increasingly popular in G protein-coupled receptor (GPCR) drug discovery for several reasons. First of all, the assays using the dyes are easy to perform and are of low cost compared to other assays. Second, most non-Gαq-coupled GPCRs can be tweaked to modulate intracellular calcium by co-transfection with promiscuous or chimeric/mutated G proteins making the calcium assays broadly applicable in GPCR research. Third, the price of instruments capable of measuring fluorescent-based calcium indicators has become significantly less making them obtainable even for academic groups. Here, we present a protocol for measuring changes in intracellular calcium levels in living mammalian cells based on the fluorescent calcium binding dye, fluo-4.
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Acknowledgments
Our work on FLIPR® assays of intracellular calcium has been supported by The Danish Medical Research Council, the Lundbeck Foundation, the Villum Kann Rasmussen Foundation, the Novo Nordisk Foundation, Fonden af 17-12-1981, the Augustinus Foundation, and the Direktør Ib Henriksen Foundation.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Hansen, K.B., Bräuner-Osborne, H. (2009). FLIPR® Assays of Intracellular Calcium in GPCR Drug Discovery. In: Leifert, W. (eds) G Protein-Coupled Receptors in Drug Discovery. Methods in Molecular Biology, vol 552. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-317-6_19
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DOI: https://doi.org/10.1007/978-1-60327-317-6_19
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