A Colorimetric Method for the Determination of Lipoxygenase Activity Suitable for Use in a High Throughput Assay Format

✍ Scribed by Neil B. Waslidge; David J. Hayes

• Publisher: Elsevier Science
• Year: 1995
• Tongue: English
• Weight: 87 KB
• Volume: 231
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.1995.0063

✦ Synopsis

throughput assay. Therefore, there is a need to develop A rapid and sensitive method for measuring the aca method that is rapid, reproducible, and suitable for tivity of lipoxygenase is described. The assay is based adaptation to a high throughput assay format. Here on the principle that under acidic conditions a lipid we describe a modification of the colorimetric assay of hydroperoxide can oxidize Fe 2/ to Fe 3/ which then oxi-Jiang et al. (1) namely the ferric oxidation of xylenol dizes xylenol orange to form a product that absorbs orange (FOX) 2 to the measurement of lipoxygenase acstrongly in the visible region (see Z-Y. Jiang, A. C. S. tivity. We used solubilized human platelet 12-LO as an Woollard, and S. P. Wolff, Lipids 26, 853-856, 1991). enzymatic source with which to characterize the assay. This methodology was modified to measure lipoxygen-We believe that our methodology is equally applicable ase activity and the system was optimized using plateto the other lipoxygenase enzymes.

let 12-lipoxygenase. This assay is suitable for use in a 96-well microtiter plate and may be utilized as a high MATERIALS AND METHODS throughput screen for the identification of novel lipoxygenase inhibitors.