*5.2. Toxins*

Venoms of *Bothrops jararaca* and *B. neuwiedi* were obtained from snakes housed in captivity at the Butantan Institute Serpentarium (Laboratório de Herpetologia). Jar and Jar-C were isolated from *B. jararaca* snake venom, and BnP1 was isolated from *B. neuwiedi* snake venom, as previously described [13,19,21]. The purity of toxins was tested using 12.5% SDS–PAGE under reducing conditions [52], and all samples were subjected to treatment with Triton X-114 to remove eventual remaining LPS contamination [53]. After Triton X-114 treatment, LPS absence was confirmed by the Limulus Amebocyte Lysate test (LAL, Charles River). Jar and BnP1 used for the LAL assay were previously heated at 70 ◦C to inactivate the proteolytic activity, and the biological activities of toxins after Triton X-114 treatment were tested by determining the hemorrhagic activity (Jar and BnP1) or by performing a platelet aggregation inhibition assay (Jar-C). All the toxins used in the experiments were LPS-free. The doses of toxins used were based in a previous study [9].
