PrP Extraction and Western Blot Analysis

PrP^Sc extraction and the following WB analyses were carried out on the brainstem samples collected at necropsy from the experimentally challenged goats. Brain tissue from healthy cattle, C-BSE and L-BSE samples, used for the inoculum, and goats previously confirmed positive or negative for classical scrapie were also analyzed as controls. The analyses were performed as previously reported^{11 (link)}. PrP^Sc was detected using SAF84 (mouse; Cayman Chemical Co.; Cat. No. 189775) monoclonal antibody and an anti-mouse antiserum conjugated with alkaline phosphatase. Reaction was revealed by a chemiluminescent substrate and visualized on Hyperfilm ECL (Amersham GE-Healthcare, Life Sciences) or by a gel documentation (UVI Prochemi, Uvitec). Image Lab^TM analysis software (Bio-Rad) was used to analyse blot images.