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A fluorimetric assay for human reticulocyte 15-lipoxygenase-1 activity
- Source :
- Analytical biochemistry. 426(1)
- Publication Year :
- 2012
-
Abstract
- 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 IC(50) 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.
- Subjects :
- Reticulocytes
Biophysics
Transfection
Biochemistry
Rhodamine 123
Fluorescence spectroscopy
chemistry.chemical_compound
Lipoxygenase
Reticulocyte
medicine
Arachidonate 15-Lipoxygenase
Humans
Fluorometry
Lipoxygenase Inhibitors
Molecular Biology
Chromatography
biology
Rhodamines
food and beverages
Cell Biology
Fluorescence
Nordihydroguaiaretic acid
medicine.anatomical_structure
HEK293 Cells
chemistry
biology.protein
Quercetin
Fisetin
Subjects
Details
- ISSN :
- 10960309
- Volume :
- 426
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Analytical biochemistry
- Accession number :
- edsair.doi.dedup.....d56749229a745e32729ffddf9b3d7cd0