pH-Insensitive FRET Voltage Dyes
Michael P. Maher*,
Nyan-Tsz Wu,
Hong Ao
Johnson & Johnson Pharmaceutical Research and Development
* To whom correspondence should be addressed. E-mail: mmaher1{at}prdus.jnj.com.
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Abstract |
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Many high-throughput ion channel assays require the use of voltage-sensitive dyes to detect channel activity in the presence of test compounds. Dye systems employing Förster resonance energy transfer (FRET) between 2 membrane-bound dyes are advantageous in combining high sensitivity, relatively fast response, and ratiometric output. The most widely used FRET voltage dye system employs a coumarin fluorescence donor whose excitation spectrum is pH dependent. The authors have validated a new class of voltage-sensitive FRET donors based on a pyrene moiety. These dyes are significantly brighter than CC2-DMPE and are not pH sensitive in the physiological range. With the new dye system, the authors demonstrate a new high-throughput assay for the acid-sensing ion channel (ASIC) family. They also introduce a novel method for absolute calibration of voltage-sensitive dyes, simultaneously determining the resting membrane potential of a cell. (Journal of Biomolecular Screening XXXX:xx-xx)
