title: Development and optimization of FLIPR high throughput calcium assays for ion channels and GPCRs.
abstract: Ca(2+) permeable ion channels and GPCRs linked to Ca(2+) release are important drug targets, with modulation of Ca(2+) signaling increasingly recognized as a valid therapeutic strategy in a range of diseases. The FLIPR is a high throughput imaging plate reader that has contributed substantially to drug discovery efforts and pharmacological characterization of receptors and ion channels coupled to Ca(2+). Now in its fourth generation, the FLIPR(TETRA) is an industry standard for high throughput Ca(2+) assays. With an increasing number of excitation LED banks and emission filter sets available; FLIPR Ca(2+) assays are becoming more versatile. This chapter describes general methods for establishing robust FLIPR Ca(2+) assays, incorporating practical aspects as well as suggestions for assay optimization, to guide the reader in the development and optimization of high throughput FLIPR assays for ion channels and GPCRs.
authors.affiliation: Institute for Molecular Bioscience, The University of Queensland, St Lucia, QLD, Australia. i.vetter@uq.edu.au
authors.forename: Irina
authors.initials: I
authors.lastname: Vetter
mesh_terms: D000818:Animals; D002118:Calcium / Q000032:analysis* / Q000378:metabolism; D005456:Fluorescent Dyes; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007473:Ion Channels / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*
chemical_list: D005456:Fluorescent Dyes; D007473:Ion Channels; D043562:Receptors, G-Protein-Coupled; D002118:Calcium