cells centrifuged, resuspended in 0.01 M Tris-HCl, pH 7.5, sonicated, centrifuged, supernatant subjected to streptomycin sulfate (10%) and ammonium sulfate (70%) precipitation, precipitated protein dissolved in same buffer, desalted on a G-25 column
recombinant cells are centrifuged, pellet washed with PBS, pH 7.4, resuspended in lysis buffer (50 mM Tris-HCl, pH 8.0), incubated, sonicated, centrifuged, Ni2+ -NTA agarose column, washed with buffer (50 mM Tris-HCl, pH 8.0), eluted imidazole, SDS-PAGE, dialyzed with 50 mM Tris-HCl, pH 8.0, storage with 20% glycerol at -80°C
cells centrifuged, resuspended in 0.01 M Tris-HCl, pH 7.5, sonicated, centrifuged, supernatant subjected to streptomycin sulfate (10%) and ammonium sulfate (70%) precipitation, precipitated protein dissolved in same buffer, desalted on a G-25 column
cells centrifuged, resuspended in 0.01 M Tris-HCl, pH 7.5, sonicated, centrifuged, supernatant subjected to streptomycin sulfate (10%) and ammonium sulfate (70%) precipitation, precipitated protein dissolved in same buffer, desalted on a G-25 column