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Analytical Techniques for Assaying Nitric Oxide Bioactivity
Published on: June 18, 2012
A nonradioactive assay for nitric oxide synthase activity in tissue extracts
Hazel Sutherland1, Olga Zolle, Roba Khundkar
1Department of Human Anatomy and Cell Biology, University of Liverpool, UK.
Methods in Molecular Biology (Clifton, N.J.)
|June 17, 2004
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.

