MMP protein profiles of human primary gastric mucosa of H. pylori–infected and uninfected donors were analyzed with the Proteome Profiler Human Protease Array Kit (ARY021B, R&D Systems) according to the manufacturer’s instructions. Briefly, an equivalent amount (about 0.3 mm3) of human primary gastric mucosa from H. pylori–infected patients and paired uninfected donors was homogenized in 0.2-ml sterile protein extraction reagent and centrifuged. Quantitation of sample protein concentration was achieved by bicinchoninic acid total protein assay. First, each membrane was incubated in an array blocking buffer for 1 hour on a rocking platform. Then, 15 μl of reconstituted protease-detecting Ab cocktail was added to each prepared sample at room temperature for 1 hour. Array blocking buffer was then removed from the wells, and the preprepared sample/Ab mixtures were added to the corresponding membrane and incubated overnight at 4°C. Each membrane was then washed three times with washing buffer before being incubated with streptavidin-HRP for 30 min at room temperature and then developed with 1 ml of the prepared chemi reagent mix for 1 min.

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