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Fig. 1 | Journal of Venomous Animals and Toxins including Tropical Diseases

Fig. 1

From: Antivenomic approach of different Crotalus durissus collilineatus venoms

Fig. 1

Antivenom recognition of C. d. collilineatus venoms and fractions performed by indirect enzyme-linked immunosorbent assay (ELISA) and chromatographic profiles of pooled venom. The 96-well plates were sensitized with 2 μg of (a) venoms (1–22) and (c) RP-FPLC fractions (0–44) diluted to 100 μL with carbonate-bicarbonate buffer (pH 9.6). The commercial anticrotalic serum (1,100) from Instituto Butantan was used to evaluate its capacity to recognize the venoms and their fractions using antihorse polyclonal antibodies peroxidase-labeled (1,3000) as secondary antibody. Positive control (C+): wells sensitized with anticrotalic antivenom (represented by horizontal dashed lines). Negative control (C-): non-sensitized wells. Absorbance reading was performed at 490 nm. Data are presented as mean ± SD, which were analyzed by ANOVA and Tukey’s multiple comparison test (quadruplicate assay). *p < 0.05, **p < 0.01 and ***p < 0.001 compared to C-; #p < 0.05, ##p < 0.01 and ###p < 0.001 compared to C+. The arrow indicates crotamine-positive venom and fractions. (b) RP-FPLC of C. d. collilineatus pooled venom (22 mg) on a C18 column was carried out in a segmented concentration gradient from 6.3 to 100% of solution B (80% ACN in 0.1% TFA, represented by the blue dashed line) at a flow rate of 5 mL/min. Inset panel – whole chromatographic profile without magnification

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