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The pellets remaining after the second centrifugation were washed with buffer-1 for 15 min and centrifuged at 15,000 g for 10 min, and the resulting pellets were incubated with buffer-2 [20 mM HEPES/KOH (pH 7.8), 1.5 mM MgCl 2 , 420 mM NaCl, 0.2 mM EDTA, 25% glycerol, 0.5 mM dithiothreitol, 0.2 mM phenylmethylsulfonyl fluoride, and 1 mM Na 3 VO 4 ] for 20 min
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The mixture was stirred for 8h at room temperature
GHK that occurs naturally in the body has a high affinity for copper and is involved in the binding and safe transport of copper throughout the body, where it is used by cells and enzymes as a cofactor
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