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Purification (Commentary)
liver is homogenized, centrifuged, supernatant mixed with 8 mM calcium chloride, microsomes separated by centrifugation, pellet resuspended in 50 mM Tris-HCl with 0.1 mM ethylendiaminetetraacetic acid and 20% glycerol, pH 7.4
cells centrifuged, washed with TD buffer (50 mM Tris-HCl, 1 mM dithiothreitol (DTT), pH 7.6), centrifuged, bacterial pellet resuspended in same buffer, sonicated, centrifuged, supernatant used as cell extract or further purified with ion-exchange chromatography with a HiTrap Q-Sepharose column, active fractions are desalted with PD-10 desalting column, concentrated with Vivaspin concentrator
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