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. 2016 Aug 25;6:32036. doi: 10.1038/srep32036

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Copyright © 2016, The Author(s)

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(A) Size-exclusion chromatography of the crude venom. The elution was monitored at 280 nm. The fractions of peak 4 (horizontal bar) were pooled and sub-fractionated on RP-HPLC. (B) RP-HPLC chromatography of peak 4. The elution was monitored at 215 nm. The inhibitory activities of the individual fractions on FX activation by the ETC were measured (dotted line). The peaks indicated by solid arrow (contains exactin) and open arrow (contains other anticoagulant proteins, which are being characterized) significantly prolonged the plasma prothrombin time (Fig. S1). (C) The re-purification of exactin on a shallow gradient of 30–36% solvent B. The elution was monitored at 215 nm. (D) The ESI-MS of exactin showing three peaks of mass/charge (m/z) ratio ranging from +4 to +6 charges. The mass of exactin was determined to be 6621.12 ± 0.22 Da (inset).