arachidonic-acid-omega-9-hydroperoxide and xylenol-orange

A rapid and sensitive method for measuring the activity of lipoxygenase is described. The assay is based on the principle that under acidic conditions a lipid hydroperoxide can oxidize Fe2+ to Fe3+ which then oxidizes xylenol orange to form a product that absorbs strongly in the visible region (see Z-Y. Jiang, A. C. S. Woollard, and S. P. Wolff, Lipids 26, 853-856, 1991). This methodology was modified to measure lipoxygenase activity and the system was optimized using platelet 12-lipoxygenase. This assay is suitable for use in a 96-well microtiter plate and may be utilized as a high throughput screen for the identification of novel lipoxygenase inhibitors.

Topics: 12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; Arachidonate 12-Lipoxygenase; Blood Platelets; Colorimetry; Ferric Compounds; Fluorescent Dyes; Humans; Hydroxyeicosatetraenoic Acids; Leukotrienes; Linear Models; Oxidation-Reduction; Phenols; Platelet Aggregation Inhibitors; Sensitivity and Specificity; Solubility; Spectrophotometry; Sulfoxides; Time Factors; Xylenes