FLIPR assays of intracellular calcium in GPCR drug discovery.

Kasper B. Hansen, Hans Bräuner-Osborne

Research output: Contribution to journalArticlepeer-review

Abstract

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
Pages (from-to)269-278
Number of pages10
JournalMethods in molecular biology (Clifton, N.J.)
Volume552
DOIs
StatePublished - 2009

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