FIG. 1.

Expression and purification of rNSP5. Proteins were denatured in sample buffer containing LDS and dithiothreitol (NuPAGE LDS sample buffer; Invitrogen) and resolved by PAGE on NuPAGE 10% Bis-Tris polyacrylamide gels. Proteins were detected by staining the gel with Coomassie blue (A). To locate rNSP5, protein in the gel was transferred onto nitrocellulose and the blot was probed sequentially with guinea pig anti-NSP5 polyclonal antisera (1:750 dilution) and goat anti-guinea pig horseradish peroxidase-conjugated antibody (1:5,000 dilution) (47). The blot was developed with the Sigma Fast system (B). Lanes: M, protein molecular size standards; 1, bacterial lysate prior to induction; 2, bacterial lysate after induction with IPTG; 3, His-tagged rNSP5 eluted from Ni affinity column. K, kilodaltons.