| Assay Method Information | |
| | Cell-Free Assay |
| Description: | The enzyme activity of 5-LOX was determined fluorescence spectrophotometrically by oxidation of the substrate H2DCFDA to the highly fluorescent 2',7'-dichlorofluorescein product during 5-LOX s catalytic reaction . The reactions were initiated by the addition of AA as substrate and then monitored by excitation at 500 nm and emission at 520 nm utilizing a multiwall fluorometer. Fluorescence signals were recorded for 5 min with a kinetics mode program. Inhibition activities were measured as described previously. All inhibition values were tested at the concentration of 91 µM. Zileuton (reported IC50 = 0.9 µM) was used as positive control. |
| Affinity data for this assay | |
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