Chemicals and stock solutions

The solvents and chemicals used in this research were of analytical grade. Acetonitrile (ACN) and formic acid (FA) were purchased from Biosolve (Valkenswaard, The Netherlands) and water was purified using a Milli-Q plus system (Millipore, Amsterdam, The Netherlands). Bovine plasma was obtained from Biowest (Amsterdam, The Netherlands). Iodoacetamide, β-mercaptoethanol, Argatroban, Ammonium bicarbonate and calcium chloride, were obtained from Sigma-Aldrich (Zwijndrecht, The Netherlands). Sequencing grade modified trypsin was purchased from Promega Benelux B.V. (Leiden, The Netherlands), and stored and handled according to the manufacturer’s instructions. The snake venom pools used in this study were from Echis ocellatus (Nigeria), Echis carinatus (India), Echis carinatus (UAE), Echis pyramidum leakeyi (Kenya), Echis coloratus (Egypt), Crotalus horridus (USA), Macrovipera lebetina (Uzbekistan), Daboia russelii (Sri Lanka), Bothrops asper (Costa Rica), Bothrops jararaca (Brazil), Lachesis muta (Brazil), Bothriechis schlegelii (Costa Rica), Calloselasma rhodostoma (captive bred, Thailand ancestry), Hypnale hypnale (Sri Lanka), Trimeresurus albolabris (Thailand), Trimeresurus stejnegeri (Malaysia), Deinagkistrodon acutus (China), Dispholidus typus (South Africa), Rhabdophis subminiatus (Hong Kong) and Oxyuranus scutellatus (Papua New Guinea). They were sourced in house from animals held in, or historical venoms stored in, the herpetarium at the Liverpool School of Tropical Medicine, UK. These species were selected based on previous reports of coagulopathic venom activity [7, 20, 23, 24]. Venoms were stored in lyophilized form at 4°C, until reconstitution in water to make 5 mg/mL stock solutions, which were then aliquoted and stored at –80°C until use. For the initial screening of all 20 snake venoms a concentration of 1 mg/mL was used, while 5 mg/mL was used for in depth elucidation of the 7 selected venoms.