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. Author manuscript; available in PMC: 2012 Apr 5.
Published in final edited form as: Toxicon. 2010 Sep 15;56(4):544–553. doi: 10.1016/j.toxicon.2010.05.012

Fig. 1.

Fig. 1

(a) Size exclusion (SE) chromatographic profile of lyophilized crude D. r. siamensis venom (cRVV). The grey-shaded areas indicate the location of factor X activator activity using chromogenic substrate S-2765. (b) The ET profile of the pooled fraction 3 with factor X activator activity from the 300SW SE HPLC column is shown in the upper right corner. The ET was performed by using a Pharmacia Biotech PhastSystem. A polyacrylamide isoelectric focusing (IEF) PhastGel with pH gradient 3–9 was generated in the first dimension. The gel was then rotated 90°, and a volume of 3 μL (2 mg/mL) of pooled fraction 3 was applied to the pH gradient gel. The gel was stained with silver nitrate. (c) SDS-PAGE analysis of venom fractions from SE HPLC column. Crude and fractionated cRVV were run on 4–12% Bis–Tris Gel under non-reducing conditions at 200 V for 50 min. The gel was stained with RapidStain. Lane 1: SeeBlue Plus2 Markers (Invitrogen™); lane 2: cRVV (20 μg); lane 3: fraction 3 (F3; 7 μg); lane 4: fraction 4 (F4; 7 μg); lane 5: fraction 5 (F5; 7 μg); lane 6: fraction 6 (F6; 7 μg); lane 7: fraction 7 (F7; 7 μg); lane 8: fraction 8 (F8; 7 μg); lane 9: fraction 9 (F9; 7 μg). (d) DEAE anion exchange HPLC profile from fraction 3 from the 300SW SE HPLC column. The grey-shaded areas indicate the location of factor X activator activity using chromogenic substrate S-2765. (e)The ET profile of purified RVV-X. A volume of 3 μL(0.5 mg/mL) of purified RVV-X (fraction 5) was applied to the pH gradient gel. The gel was stained with silver nitrate. (f) SDS-PAGE analysis of purified RVV-X from purification step. Samples from each purification step were run on 4–12% Bis–Tris Gel under non-reducing and reducing conditions at 200 V for 50 min. Gel was stained with RapidStain. Lane 1: SeeBlue Plus2 Markers (Invitrogen™); lane 2: cRVV (20 μg); lane 3: fraction 3 from SE (6 μg); lane 4: fraction 5 from DEAE anion exchange (purified RVV-X; 6 μg); lane 5: reduced form of purified RVV-X (7 μg).