Research reportA simple chemiluminescence assay for the determination of reactive oxygen species produced by human neutrophils
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2015, Cancer LettersCitation Excerpt :The level of nitrite, the stable derivative of NO, in cell culture supernatants, and the activity of NOS in cell lysates were measured spectrophotometrically as described earlier [17]. NADPH oxidase activity was evaluated by a chemiluminescence-based assay [24]. The results were expressed as RLU/mg cell proteins.
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2012, Journal of LuminescenceCitation Excerpt :When silver nanoparticles were used as the catalysts, the formation of active oxygen-containing reactant intermediates were frequently reported [39]. These reactive oxygen species (such as OH, O2−, 1O2) in the luminol CL reaction system can react quickly with luminol in alkaline solution to emit light, as the hydroperoxide intermediate of luminol decomposes into aminophthalate [45–47]. For validating this, the following experiments were done.
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2009, Analytica Chimica ActaCitation Excerpt :Since luminol-dependent chemiluminescence reaction is a peroxidase (MPO)-requiring reaction, the availability of released cellular myeloperoxidase is a limiting factor for the chemiluminescence response. Thus, extra peroxidase, such as horseradish peroxidase, could be added to increase the sensitivity of the measuring system [23,73,76,81,83]. The reaction of luminol with HO is controversial since some authors reported that luminol-enhanced chemiluminescence may be due to the generation of HO[68,83,84], while others do not attribute the increase in luminol signal to that radical [49,69].
HaCaT keratinocytes overexpressing the S100 proteins S100A8 and S100A9 show increased NADPH oxidase and NF-κB activities
2007, Journal of Investigative DermatologyCitation Excerpt :Expression of the other transfected genes was shown by Taqman analysis using a gene-specific forward primer and a reverse primer directed against either the bovine growth hormone polyadenylation sequence (for pcDNA3.1 and pcDNA3.0 expression plasmid) or the SV40 polyadenylation sequence (for pCMV-SPORT6 expression plasmid). Superoxide production by the indicated cells was determined by a quantitative NBT assay or by an isoluminol-amplified chemiluminescence method as described previously (Liu et al., 1996). Briefly, for the NBT test, the cells (0.5–2 × 106) were incubated in phosphate-buffered saline (PBS) containing 2 mg/ml NBT (Roth, Karlsruhe, Germany) with or without 100 ng/ml PMA (Sigma-Aldrich, München, Germany) for 1 hour at 37°C.