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| | Activity Assay |
| Description: | Nox2 activity assay for primary screening: Plasma membranes were prepared from human neutrophils, which express large amounts of Nox2, stored at −80 C until use. See Curnutte et al., J. Biol. Chem. 1987, 262, 6450-6452, hereby incorporated by reference. Membranes are mixed with recombinant, purified cytosolic regulatory proteins (p67Np47N chimera, and a constitutively active mutant of Rac, Rac1(Q61)) and FAD, along with varying concentrations of compound, and LO12, which emits light when it reacts with ROS. The reaction is initiated by addition of NADPH and SDS (an artificial activator of Nox2).Nox2 assay in human neutrophils: Human neutrophils were obtained from healthy volunteers (REF). LO12 luminescence was quantified as above. Nox4 assay using stably transfected HEK cell: HEK cells stably transfected with human Nox4 were incubated with varying concentrations of compound along with LO12. |
| Affinity data for this assay | |
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