Purification of recombinant Tpr from SF9 cells and C-Tpr (residues 1626-2348) from bacteria. Panel A: Coomassie stained SDS-PAGE (8%) of a representative 6×His-Tpr purification. Shown are the cell lysate following centrifugation, washes (800 mM NaCl, 20 mM HEPES pH7.5, 10% glycerol, 15 mM imidazole and protease inhibitors) to remove material non-specifically bound to the resin and seven elution fractions (100 mM NaCl, 20 mM HEPES, 150 mM imidazole and 5 mM beta-mercaptoethanol with protease inhibitors). Panel B: Western analysis of the samples in Fig.1 Panel A, probed with a combination of antibodies to the N-terminus and the C-terminus of Tpr. Panel C: Coomassie stained SDS-PAGE (8%) of a representative purified CTpr(1626-2348).
Ben-Efraim et al. BMC Cell Biology 2009 10:74 doi:10.1186/1471-2121-10-74