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The complex was concentrated to 0.5 ml using Ultra Centrifugal Filter (ThermoFisher MWCO, 100 kDa) and subjected to size-exclusion chromatography on a Superdex 200 Increase 10/300 column (GE Healthcare) that was pre-equilibrated with 20 mM HEPES pH 7.5, 100 mM NaCl, 2 mM MgCl 2 , 5 M ligands, 0.00075%(w/v) LMNG, 000025% glyco-diosgenin (GDN, Anatrace) and 0.00015% (w/v) CHS to separate complex from contaminants

[DOI] [PubMed] [Google Scholar] 73.Avila MA, Corrales FJ, Ruiz F, Snchez-Gngora E, Mingorance J, Carretero MV, Mato IM

Determination of Zn 2+ -to-protein stoichiometry The stoichiometry of Zn 2+ complexes obtained after reconstitution and purification of holo-forms was determined using two different approaches

10.1111/j.1469-7580.2007.00847.x Roche AF, Wainer H, Thissen D: The Knee Joint as a Biological Indicator