Purification of recombinant His6-Gtr1 wild-type protein by immobilized metal affinity chromatography. Coomassie Brilliant Blue stained 12 % (w/v) acrylamide SDS gel containing following samples: M, prestained protein molecular weight marker (kDa); lane 1, aliquot of sonicated total cell lysate; lane 2, aliquot of soluble fraction of total cell lysate after centrifugation at 40,000 g; lane 3, flow-through protein from the Ni2+-NTA column, and lanes 4–9, eluted fractions of His6-Gtr1 wild-type protein. Prior to the GTPase assay, an aliquot of eluted His6- Gtr1 wild-type protein from the Ni2+-NTA column was verified by Western blot analysis (right panel) using anti-His antibody.
Sengottaiyan et al. BMC Biochemistry 2012 13:11 doi:10.1186/1471-2091-13-11