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  • Solubilised proteins were concentrated using a 5000 molecular-weight cutoff membrane (Centricon, Millipore), diluted 1∶1 in 20 mM Tris (pH 8.0), 5 mM imidazole, 150 mM NaCl, 5 mM DTT and purified by nickel-chelate affinity chromatography (Ni-Sepharose, GE Healthcare) in an AKTA FPLC system (GE Healthcare).
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