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. 2011 Oct;10(10):1357–1366. doi: 10.1128/EC.05041-11

Fig. 1.

Fig. 1.

Copurification of Ncb2 with NRE-binding proteins. (A) Schematic representation of purification procedure. (B) Total cell extract from strain SC5314 was fractionated by ammonium sulfate, and the 50 to 60% fraction enriched in NRE-binding activity was loaded on a Q-Sepharose column. All fractions eluted were tested for NRE-binding activity by EMSA and were analyzed by 12% SDS-PAGE, followed by Coomassie staining. The left panel shows a representative picture of the NRE-binding activity of Q-Sepharose column eluates, and their respective protein profiles are shown in the right panel. (C) The left panel shows the NRE-binding activity of heparin sulfate column eluates. A silver-stained 12% acrylamide gel showing the protein profiles of the respective fractions is shown on the right. (D) Fractions from a heparin-agarose column were subjected to Sephacryl S-100 gel filtration chromatography, and the fractions obtained were analyzed by EMSA. The NRE-binding fractions eluted immediately after the void volume was pooled and was subjected to 12% SDS-PAGE and analyzed by silver staining.