Fig. 1

Purification of BaltDC.(a) Anion-exchange chromatography of B. alternatus crude venom on a DEAE-Sephacel column (2.5 × 20 cm) equilibrated with 0.05 M ammonium bicarbonate (pH 7.8) and eluted with a convex concentration gradient of the same buffer (0.05–1 M). (b) Gel filtration on Sephadex G-75 column (1.0 × 100 cm): ninth peak was applied to the column and eluted with 0.05 M ammonium bicarbonate. Fractions of 3.0 mL/tube were collected and the absorbance was read at 280 nm. (c) SDS-PAGE: Lane 1 – standard proteins; lane 2 – BaltDC under non-reducing conditions; lane 3 – BaltDC under reducing conditions. The molecular mass standard proteins used were phosphorylase b (97 kDa), bovine serum albumin (66 kDa), ovalbumin (45 kDa), carbonic anhydrase (30 kDa) and soybean trypsin inhibitor (20.1 kDa). Gels were stained with Coomassie blue R-250, 0.2%. (d) Reverse-phase HPLC on a Source 15RPC ST column (4.6 × 100 mm) equilibrated with 0.1% trifluoroacetic acid (TFA) and eluted with a linear concentration gradient from 0 to 100% of solution B (70% acetonitrile in 0.1% TFA)