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The resin was then collected by centrifugation at 500 g for 10 min and washed in gravity flow column (Sangon Biotech) with five column volumes of buffer containing 20 mM HEPES (pH 7.5), 100 mM NaCl, 10% (v/v) glycerol, 5 mM MgCl2, 1 mM MnCl2, 25 M TCEP, 0.1% (w/v) LMNG, 0.02% (w/v) CHS, 2 M GLP-1 or LY3502970 and/or 10 M compound 2, followed by washing with fifteen column volumes of buffer containing 20 mM HEPES (pH 7.5), 100 mM NaCl, 10% (v/v) glycerol, 5 mM MgCl2, 1 mM MnCl2, 25 M TCEP, 0.03% (w/v) LMNG, 0.01% (w/v) glyco-diosgenin (GDN, Anatrace), 0.008% (w/v) CHS, 2 M GLP-1 or LY3502970 and/or 10 M compound 2

10.1016/j.jbc.2022.101796 8 AldahhanR