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J Biomol Screen. 2007 Aug;12(5):656-67. Epub 2007 May 21.

pH-Insensitive FRET voltage dyes.

Author information

1
Johnson & Johnson Pharmaceutical Research and Development L.L.C., San Diego, California 92121, USA. mmaher1@prdus.jnj.com

Abstract

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.

PMID:
17517905
DOI:
10.1177/1087057107302113
[Indexed for MEDLINE]

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