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FluxOR™, a Universal Potassium Channel Assay Screening BacMam Delivered and Stably Expressed Targets
Daniel W. Beacham1, Kelly J. Cassutt3, Michael O’Grady1,David Thompson2, David Piper2 and Kyle Gee1
1Invitrogen Corporation • 29851 Willow Creek Rd • Eugene, Oregon 97402 • USA
2Invitrogen Corporation • 501 Charmany Dr • Madison, Wisconsin 53719 • USA
3Hamamatsu Photonics • 360 Foothill Rd • Bridgewater, New Jersey 08807 • USA
Sponsored by Invitrogen Corporation, part of Life Technologies
Published in December 2009 (p.6) DOI:  10.2144/000113329
Potassium ion channels are important targets in drug discovery and safety testing for which current assays suffer expense, low throughput or lack of proximity to the target. A robust and homogenous assay that allows for functional HTS measurements of potassium channel activity would give the screening community a useful tool for the interrogation of potassium channels in compound library screening. Invitrogen’s Molecular Probes Labeling and Detection Technologies has developed an equilibrium based assay for HTS measurement of potassium ion channel activity called the FluxOR™ Thallium Assay Kit. Using thallium influx as a surrogate indicator of potassium ion channel activity, the assay is based on the activation of FluxORTM, a novel fluorescent dye with a high affinity for thallium (Figure 1) that reports channel activity with a large fluorogenic response that is proportional to the number of open potassium channels on the cell. In contrast to competing thallium flux methods or published applications with BTC, (Weaver et al, Hoegaard et al), FluxORTM dye requires lower thallium concentrations for a larger signal window and normal chloride conditions. Combined with BacMam Target delivery (Kost et al, Ames et al), many potassium channels, including hERG, are now amenable to high throughput screening in a variety of cellular backgrounds, including primary human aortic smooth muscle, allowing flexibility of genetic context for maximum biological relevance.
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Cell and tissue culture-stem cells