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. 2008 Nov 11;41(Pt 6):1150–1160. doi: 10.1107/S0021889808033797

Figure 3.

Figure 3

Expression, purification and characterization of the full-length AfCopB and its N-terminal fragment AfCopB-N. (a) SDS–PAGE gel following the purification procedure of the full-length AfCopB. Protein samples were run on a 4–12% bis-tris gel in a 3-(N-morpholino)propanesulfonic acid (MOPS) buffer. The lane labeled with WC is for the whole cell lysate, M is the crude membrane, Ni is the eluent from the Ni-NTA column and SEC is the purified AfCopB after size exclusion chromatography. The two major bands after Ni-NTA chromatography are the full-length AfCopB and a co-purified N-terminal fragment of AfCopB named AfCopB-N. (b) Elution profile of a full-length AfCopB sample eluted from SEC. The blue line represents the profile for the protein sample from a Superdex 200 column, showing two peaks. The first one is from the full-length AfCopB and the second from the co-purified AfCopB-N fragment. The pink line is from commercial molecular weight standards. (c) SDS–PAGE gel following the purification of AfCopB-N. Protein sample was run on a 12% bis-tris gel in MOPS buffer. The lane labeled with WC is for the whole cell lysate, M is crude membrane, Ni is eluent from the Ni-NTA column and SEC is the purified AfCopB302 after SEC. (d) The SEC profile of AfCopB-N after a Superdex 200 column.