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. 2012 Jun;31(3):196–202. doi: 10.1089/hyb.2012.0005

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Copyright 2012, Mary Ann Liebert, Inc.

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FIG. 1. — Expression and purification of UbcH10. (A) SDS-PAGE analysis of the recombinant UbcH10 protein. Expressed proteins were analyzed by 12% SDS-PAGE stained with Coomassie Bright Blue. Lanes 1 and 2 represent total cell lysate (TCL) after solubilization under the denatured condition from Escherichia coli BL21, uninduced and induced with 1.0 mM IPTG, respectively. Recombinant UbcH10 protein is indicated by arrow. Lanes 3 and 4 represent soluble supernatant and insoluble pellet extracts from E. coli BL21 cell lysates, respectively. Lane 5 was the soluble recombinant Ubch10 protein purified by Ni-NTA agarose. Purified recombinant UbcH10 protein about 20 kDa is indicated by arrow. M, protein molecular mass standards. (B) Identification by Western blot analysis using anti-His polyclonal antibody. M, protein marker (17 kDa); lane 1, purified recombinant UbcH10 protein; lane 2, supernatant of BL21 cell lysate with pET-32a(+).