| Assay Method Information | |
| | NOX2 Inhibition Assay |
| Description: | Evaluation of the NOX4 inhibitors on NOX2 activity is assessed in human blood-derived granulocytes.Different concentrations of the test compounds (30 nM-100 μM), DMSO (as a negative control) or diphenyleneiodonium chloride (DPI; 30 μM, as a positive control), prepared in assay buffer (PBS containing CaCl2 and MgCl2+0.1% bovine serum albumin (BSA)), are placed in 384 well plates. Afterwards, freshly isolated granulocytes are added to the wells containing the compounds in assay buffer and incubated for 1 h at 37° C. Next, a solution of N-formylmethionine-leucyl-phenylalanine (fMLP; 730 nM), phorbol myristate acetate (PMA, 100 nM) (stimuli of ROS production) and L-012 (200 uM, ROS indicator) is added to the wells and the cells are incubated for additional 30 mins at 37° C. Afterwards luminescence is measured in a SpectraMax M5 microplate reader. |
| Affinity data for this assay | |
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