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One-step purification of SAICAR synthase by affinity chromatography. The cell lysate was incubated with Ni–NTA resin at 277 K for 2 h. The resin was then collected and washed with wash buffer containing an imidazole gradient (see §2 for details). The SDS–PAGE profile is shown. Lane 1, protein molecular-weight markers (kDa); lane 2, 10 mM imidazole eluant; lane 3, 20 mM imidazole eluant; lane 4, 50 mM imidazole eluant; lane 5, 200 mM imidazole eluant.
