Development and optimization of FLIPR high throughput calcium assays for ion channels and GPCRs.

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.