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On the use of L-012, a luminol-based chemiluminescent probe, for detecting superoxide and identifying inhibitors of NADPH oxidase: a reevaluation

Free Radical Biology and Medicine, ISSN: 0891-5849, Vol: 65, Page: 1310-1314
2013
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L-012, a luminol-based chemiluminescent (CL) probe, is widely used in vitro and in vivo to detect NADPH oxidase (Nox)-derived superoxide (O 2 − ) and identify Nox inhibitors. Yet understanding of the free radical chemistry of the L-012 probe is still lacking. We report that peroxidase and H 2 O 2 induce superoxide dismutase (SOD)-sensitive, L-012-derived CL in the presence of oxygen. O 2 − alone does not react with L-012 to emit luminescence. Self-generated O 2 − during oxidation of L-012 and luminol analogs artifactually induce CL inhibitable by SOD. These aspects make assays based on luminol analogs less than ideal for specific detection and identification of O 2 − and NOX inhibitors.

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