FLIPR assays of intracellular calcium in GPCR drug discovery

Research output: Chapter in Book/Report/Conference proceedingBook chapterResearch

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-Galpha(q)-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.
Original languageEnglish
Title of host publicationG protein-coupled receptors in drug discovery
EditorsWayne R Leifert
Place of PublicationUnited States
PublisherHumana Press
Publication date2009
ISBN (Print)978-1-60327-316-9
ISBN (Electronic)978-1-60327-317-6
Publication statusPublished - 2009
SeriesMethods in Molecular Biology

Bibliographical note

Keywords: Intracellular calcium measurement, G protein-coupled receptors, probenecid, FLIPR, FlexStation, NOVO star.

Chapter 19

ID: 13063072