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. 2013 Oct 15;5(10):1780–1798. doi: 10.3390/toxins5101780

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© 2013 by the authors; licensee MDPI, Basel, Switzerland.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).

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Reactivity of purified rabbit anti BlK-PLA₂ IgG against BlD- and BlK-PLA₂ isoforms. (A) reduced SDS-PAGE (15% gel) of: 1, molecular mass markers; 2, crude venom (20 µg); 3, BlK-PLA₂ (5 µg); 4, BlD-PLA₂ (5 µg); (B) immunoblotting of anti BlK-PLA₂ IgG against crude venom (1), BlK-PLA₂ (2) and BlD-PLA₂ (3). (C) Reactivity ofanti BlK-PLA₂ IgG against several snake venoms examined by ELISA. 96-well microtitration plates were precoated with 0.5 µg/mL of Bl-PLA₂s (pool), Bothrops species, L. muta, C. d. terrificus and Micrurus lemniscatus. Anti BlK-PLA₂ IgG was added at different dilutions. Binding was visualized by incubation with peroxidase-coupled anti-rabbit IgG (diluted 1:12,000) and subsequent peroxidase-catalyzed conversion of O-phenylenediamine (OPD). The absorbance of pre-immune serum (control) was subtracted. Data shown represent the average of two independent experiments, with error bars indicating the maximum and minimum deviation from the average.