文献类型：期刊文献

英文题名：A fluorimetric assay for human reticulocyte 15-lipoxygenase-1 activity

作者：Lu, Weiqiang[1];Zhao, Xue[1];Zou, Shien[2];Huang, Jin[1]

机构：[1]E China Univ Sci & Technol, Shanghai Key Lab New Drug Design, Shanghai Key Lab Chem Biol, Sch Pharm, Shanghai 200237, Peoples R China;[2]Fudan Univ, Obstet & Gynecol Hosp, Shanghai 200011, Peoples R China

年份：2012

卷号：426

期号：1

起止页码：66

外文期刊名：ANALYTICAL BIOCHEMISTRY

收录：;EI(收录号：20201908624074);WOS:【SCI-EXPANDED(收录号:WOS:000305045000012)】；

基金：This work was supported by the 111 Project (Grant B07023), the National Natural Science Foundation of China (Grants 10979072, 81102420), the Innovation Program of the Shanghai Municipal Education Commission (Grant 10ZZ41), the Specialized Research Fund for the Doctoral Program of Higher Education of China (Grant 20090074120012), and the Shanghai Committee of Science and Technology (Grant 11DZ2260600). The authors thank Professor Colin D. Funk (Department of Biochemistry, Queen's University, Canada) for the plasmid pCDNA3-15-LOX-1.

语种：英文

外文关键词：15-Lipoxygenase-1; Inhibitor; Fluorimetric assay; Dihydrorhodamine 123

摘要：A rapid and sensitive fluorescence-based assay for the determination of human 15-lipoxygenase-1 (15-LOX-1) activity is described in this article. The assay utilizes the ability of 15-LOX-1-generated lipid hydroperoxides to oxidize nonfluorescent dihydrorhodamine 123, producing the highly fluorescent dye rhodamine 123. Formation of rhodamine 123 can be monitored through fluorescence spectroscopy using Ex/Em of 500 nm/536 nm. The IC50 values of three well-known 15-LOX-1 inhibitors, nordihydroguaiaretic acid, quercetin, and fisetin, were evaluated in 96- and 384-well formats, and they conform to previously reported data. We believe this assay can be broadly used for the discovery of novel lipoxygenase inhibitors. (C) 2012 Elsevier Inc. All rights reserved.