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Cell pellets were then lysed using EmulsiFlex-C3 in a buffer containing 100 mM HEPES-KOH pH 7.5, 300 mM KCl, 2 mM dithiothreitol (DTT), 5 mM MgCl 2 , 5 mM imidazole, 10% glycerol, 0.1% IGEPAL CA-630 and protease inhibitor cocktail

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Am J Respir Crit Care Med 171 : 431–439 Fitzpatrick AM, Brown LA, Holguin F, Teague WG, National Institutes of Health/National Heart, Lung, and Blood Institute Severe Asthma Research Program 2009 Levels of nitric oxide oxidation products are increased in the epithelial lining fluid of children with persistent asthma