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The resin was washed with 15 column volumes of Wash Buffer 1 containing 20 mM HEPES pH 7.5, 100 mM NaCl, 2 mM MgCl 2 , 15 mM imidazole, 0.1% LMNG, 0.02% CHS, 10 μM ligands (ghrelin or GHRP-6) and 15 column volumes of Wash Buffer 2 containing 20 mM HEPES, pH 7.5, 100 mM NaCl, 2 mM MgCl 2 , 25 mM imidazole, 0.01% LMNG, 0.005% GDN (Anatrace), 0.003% CHS, 10 μM ligands (ghrelin or GHRP-6)