title: A nonradioactive assay for nitric oxide synthase activity in tissue extracts.
abstract: 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.
authors.affiliation: Department of Human Anatomy and Cell Biology, University of Liverpool, UK.
authors.forename: Hazel
authors.initials: H
authors.lastname: Sutherland
authors.forename: Olga
authors.initials: O
authors.lastname: Zolle
authors.forename: Roba
authors.initials: R
authors.lastname: Khundkar
authors.forename: Alec W M
authors.initials: AW
authors.lastname: Simpson
authors.forename: Jonathan C
authors.initials: JC
authors.lastname: Jarvis
authors.forename: Stanley
authors.initials: S
authors.lastname: Salmons
mesh_terms: D000818:Animals; D002956:Citrulline / Q000737:chemistry; D019793:Fluorescein / Q000737:chemistry*; D018482:Muscle, Skeletal / Q000201:enzymology*; D009569:Nitric Oxide / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D013050:Spectrometry, Fluorescence / Q000379:methods; D014020:Tissue Extracts / Q000378:metabolism*
chemical_list: C111205:4,5-diaminofluorescein; D014020:Tissue Extracts; D002956:Citrulline; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D019793:Fluorescein