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In 1818, Thnard had already observed that the hydrogen peroxide was decomposed by animal tissues, with oxygen discharge

The plates were then washed twice with ice-cold PBS, scraped, pelleted 250 g at 4 C for 3 min and lysed with RIPA buffer (50 mM Tris-Cl (pH = 8), 150 mM NaCl, 5 mM EDTA, 1 % NP-40, 0.5 % sodium deoxycholate, 0.1% SDS, 1x protease and 1x phosphatase inhibitors)

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Lysyl oxidase, the copper-dependent enzyme that catalyzes the oxidative deamination of lysine residues in collagen and elastin (a step required for the cross-linking that gives mature collagen and elastin their mechanical properties), requires copper as a cofactor

doi: 10.1038/s41598-017-01732-1 80 ConnellyPJPrenticeNPCouslandGBonhamJ