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We describe a fluorescence assay for nitric oxide synthase activity based on a new indicator, 4,5-diaminofluorescein (DAF-2). The method offers the advantage of being safer and more convenient than the citrulline radioassay in common use. The rapid and irreversible binding of DAF-2 to oxidized nitric oxide (NO) enables NO production to be measured in real time. The protocol is applied to the measurement of nitric oxide synthase in crude extracts of skeletal muscle.

Original publication

DOI

10.1385/1-59259-807-2:105

Type

Journal article

Journal

Methods mol biol

Publication Date

2004

Volume

279

Pages

105 - 111

Keywords

Animals, Citrulline, Fluorescein, Muscle, Skeletal, Nitric Oxide, Nitric Oxide Synthase, Spectrometry, Fluorescence, Tissue Extracts