| Solution Information | help | |
| Enzyme: | NADPH oxidase 1 | |
| inhibitor: | BDBM67506 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay Overview: The purpose of this cell-free assay is to evaluate the ability of compounds identified as active in previous assays (the "Luminescence-based cell-based dose response assay to identify inhibitors of NADPH oxidase 1 (NOX1): Cherry picks 2" (AID 434997) assay or the "Late-stage luminescence-based cell-based dose response assay to identify inhibitors of NADPH oxidase 1 (NOX1): Purchased analogs" (AID 2808) assay) to inhibit ROS production by another cellular source, xanthine oxidase. This chemiluminescence assay employs a luminol probe to monitor ROS production. Protocol Summary: In this assay, xanthine oxidase (0.5 mg/mL) was dispensed to all wells of a 96-well plate followed by test or control compounds. Hypoxanthine was dispensed to all wells, followed by the addition of horserasish peroxide and luminal and reading luminescence. The interaction of luminol with xanthine oxidase-generated ROS/superoxide inside cells yields an unstable endoperoxide that generates ligh | |
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