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. 2011 Jun 7;7(6):729–739. doi: 10.7150/ijbs.7.729

Fig 1.

Fig 1

Purification of Gigantoxin-4. (A) Cation-exchange chromatography on Hitrap Capto S (0.7 × 2.5 cm, GE, USA) using an ÄKTA purifier 100 System (GE, USA). 1 ml crude extract was applied to the column, which was eluted with a gradient of 11%, 21%, 30% and 100% NaCl (1.0 M) in 50 mM ammonium acetate buffer (pH 5.6) at a flow rate of 1 ml/min. Peak 1 and 2 show the high hemolytic activity collected. (B) Gel-filtration chromatography. 2 ml of the concentrated peak 1 protein solution was applied to the Superdex 75 column (1.6 × 60 cm, GE, USA), which was eluted with 150 mM ammonium acetate buffer (pH 5.1) with a flow rate of 1 ml/min. (C) Gel-filtration chromatography. A concentrated sample of peak 2 was applied to the Superdex 75 column. (D) SDS-PAGE analysis of mucus exudates (V1) and crude extract (V2) from S. gigantea. (E) SDS-PAGE analysis of fractions collected after Gel-filtration chromatography. (M) Marker, (1') peak 1', (2') peak 2'.