Apocynin, NADPH-oxidase inhibitor

3 product images

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  Chemical Structure - Apocynin, NADPH-oxidase inhibitor (ab120615)

  2D chemical structure image of ab120615, Apocynin, NADPH-oxidase inhibitor

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  Immunocytochemistry/ Immunofluorescence - Apocynin, NADPH-oxidase inhibitor (ab120615)

  Anti-Glutathione antibody [D8] ab19534 staining glutathione in A549 cells treated with apocynin (ab120615), by ICC/IF. Increase in glutathione expression correlates with increased concentration of apocynin, as described in literature.
  The cells were incubated at 37°C for 24h in media containing different concentrations of ab120615 (apocynin) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with Anti-Glutathione antibody [D8] ab19534 (10 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-mouse polyclonal antibody (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/250 dilution was used as the secondary antibody.
  

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  Image from Wang T et al., PloS one., 9(3): e91063. Fig 6.; doi: 10.1371/journal.pone.0091063 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

  Functional Studies - Apocynin, NADPH-oxidase inhibitor (ab120615)

  A–B. Endothelial cells were transfected with Nef cDNA, incubated for further 6 hours and treated with apocynin (200 nM), trolox (200 nM), Nox2 inhibitor (1 uM) or IKKi (100 nM). After additional 18 hours supernatants were analyzed for Nef-induced MCP-1 production (A) and endothelial cells for apoptosis using TUNEL (B). C–D. Endothelial cells were cocultured with Nef-transfected Jurkat cells for 24 h, and then treated with apocynin (200 nM), trolox (200 nM), Nox2 inhibitor (1 uM) or IKKi (100 nM) and incubated an additional 18 h, then analyzed for Nef-induced MCP-1 production (C) and apoptosis of endothelial cells (D). Data were expressed as fold MCP-1 production and apoptosis, normalized to the mean of control measurements. Data represent mean±SD from 3 separate experiments in which measurements were made in triplicate. *P<0.05, and **P<0.01.
  

  Wang T et al., PloS one., 9(3): e91063. Fig 6.; doi: 10.1371/journal.pone.0091063