Journal: Prion

Article Title: Identification of H-type BSE in Portugal

doi: 10.1080/19336896.2014.997615

Figure Lengend Snippet: Phenotypical features of the first Portuguese H-BSE. (a) PrPres immunolabelling in the reticular formation of the brainstem: linear pattern (arrow) and the predominant intraglial pattern as single granules in the white matter (arrowheads) (b). The box in picture (a) outlines the area shown in picture (b) at higher magnification. Specific PrPres signal was visualized with Vectastain–peroxidase, DAB (Vector). All sections were counterstained with Mayer's haematoxylin. The magnifications used are indicated. (c) Western blot analysis of PrPres after proteinase K treatment using Sha31 and P4 immunolabelling: Sha31 shows a higher molecular mass migration (orange arrow) by comparison to C-BSE; with P4 there is immunolabelling in the Portuguese H-BSE (PT H-BSE) similar to that presented by the H-BSE control. Absence of signal in C-BSE and L-BSE. H-BSE, C-BSE and L-BSE controls from AHVLA proficiency testing. Scrapie represents classical scrapie control. M, molecular mass marker (kDa).

Article Snippet: The confirmation of the H-type BSE phenotype was carried out using the discriminatory molecular method with the C-terminal anti-PrP antibody Sha31 and the N- terminal anti-PrP monoclonal antibody P4 (aa 101–107 bovine PrP protein, 15 R-Biopharm, 0.1mg/ml) diluted 1/500 as recommended by the European Union Reference Laboratory (AHVLA Bio-Rad TeSeE- based Hybrid Western Blot method).

Techniques: Plasmid Preparation, Western Blot, Migration, Marker