| Solution Information | help | |
| Enzyme: | Cytochrome b-245 heavy chain | |
| inhibitor: | BDBM67509 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay Overview: The purpose of this cell-based 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 NOX-2, -3, or -4 activity in a HEK/293 transfection format. This chemiluminescence assay employs a luminol probe to monitor intracellular ROS in HEK/293 cells. Protocol Summary: In the family selectivity assays, HEK/293 cells seeded into 6-well plates were cotransfected with the appropriate expression vectors for each NOX subtype. For the NOX2 assay (Assay 1), cells were transfected with pRK5-Myc- Nox2, pRK5-p67phox, pRK5-p47phox and pRK5-myc-Rac1CA-Q61L. For the NOX-3 assay (Assay 2), cells were transfected with pRK5-Myc-NOX3, pRK5-NOXO1, pRK5-NO | |
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