Improved detection of nitric oxide radical (NO.) production in an activated macrophage culture with a radical scavenger, carboxy PTIO and Griess reagent.
An improved method for the detection of nitric oxide radicals (NO. in cultures of activated macrophages was developed, involving a nitric oxide radical scavenger, 2-(4-carboxyphenyl)-4,4,5,5- tetramethylimidazoline-3-oxide-1-oxyl (carboxy PTIO) and Griess reagent. A murine macrophage-like cell line, J774.1, was activated with interferon-gamma (IFN-gamma) and bacterial lipopolysaccharide (LPS), which induced the production and secretion of NO2- into the culture supernatant. Addition of carboxy PTIO to the activated macrophages increased the amount of NO2- to 4- to 5-fold without cell damages, probably because carboxy PTIO rapidly reacted with NO. to form NO2-, which was finally assayed by the Griess reaction.
Amano
F
F
Department of Biochemistry and Cell Biology, National Institute of Health, Tokyo, Japan.
Noda
T
T
D000594:Amino Acid Oxidoreductases / Q000737:chemistry; D000818:Animals; D001565:Benzoates; D002460:Cell Line; D003497:Cyclic N-Oxides / Q000737:chemistry*; D004305:Dose-Response Relationship, Drug; D005029:Ethylenediamines; D016166:Free Radical Scavengers / Q000737:chemistry*; D005609:Free Radicals; D007093:Imidazoles / Q000737:chemistry*; D007202:Indicators and Reagents; D008262:Macrophage Activation; D008264:Macrophages / Q000737:chemistry*; D051379:Mice; D009569:Nitric Oxide / Q000096:biosynthesis / Q000737:chemistry*; D019001:Nitric Oxide Synthase; D013424:Sulfanilamides
D001565:Benzoates; D003497:Cyclic N-Oxides; D005029:Ethylenediamines; D016166:Free Radical Scavengers; D005609:Free Radicals; C095000:Griess reagent; D007093:Imidazoles; D007202:Indicators and Reagents; D013424:Sulfanilamides; C079393:1,3-dihydroxy-4,4,5,5-tetramethyl-2-(4-carboxyphenyl)tetrahydroimidazole; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D000594:Amino Acid Oxidoreductases