| Assay Method Information | |
| | NOX4 Inhibition Assay |
| Description: | NOX4 inhibition is assessed utilizing HEK293 cells stably overexpressing human NOX4 (hNOX4), generating constitutively high levels of hydrogen peroxide (H2O2). Cells are cultured in the Dulbecco's Modified Eagle Medium (DMEM) media containing 4.5 g/L glucose supplemented with 10% fetal calf serum and 250 μg/ml Zeocin in an incubator at 37° C. with 5% CO2. For the assay, cells are seeded in 384 well plates. After 24 h cells are washed and treated with several concentrations (10 nM-30 μM) of the test compounds (diluted in DMSO) or 100 μM diphenyleneiodonium chloride (DPI, used as positive control) in assay buffer consisting of phosphate-buffered saline (PBS) with 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), and incubated for 2 hours at 24° C. in a humidified incubator. |
| Affinity data for this assay | |
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