*4.5. Immunoblotting*

The purified protein (10 μg) of *T. maculosa* venom was analyzed by 12% SDS-PAGE under non-reducing conditions. Venom from *T. nattereri* (V*Tn*) was used as a control.

After SDS-PAGE and transfer to nitrocellulose membrane (pore size = 0.2 μm, Schleicher and Schüll, Dassel, Germany), the toxin was detected using plasma from mice sensitized with *T. nattereri* venom (1:20 dilution) or from Nattectin-sensitized mice followed by Goat anti-mouse IgG HRP (sc-2005 Santa Cruz, at 1:2000) as a secondary antibody (Ab).
