1.14.13.29 - 438903, 438908 1.14.13.29 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 698597 1.14.13.29 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 695586 1.14.13.29 recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain Rosetta(DE3)/pLysS by nickel affinity chromatography, gel filtration, and ultrafiltration 763974