A Comparative Study of Immunohistochemical Methods for Detecting Abnormal Prion Protein with Monoclonal and Polyclonal Antibodies

Distinct profiles of PrPd immunoreactivity in the brain of scrapie- and BSE-infected sheep: implications for differential cell targeting and PrP processing

“…Brains of sheep with scrapie show marked variation in the morphology of PrP d deposits (Miller et al, 1993;van Keulen et al, 1995;Foster et al, 1996;Hardt et al, 2000;Ryder et al, 2001). We have recently indicated that these different PrP d types and patterns can be used to obtain PrP d profiles, which appear to be mainly influenced by the source of scrapie agent and very little, if at all, by the host breed and PrP genotype (González et al, 2002).…”

“…The brains of the sheep were fixed, trimmed and embedded following standard procedures, and subjected to IHC labelling for PrP d according to previously described protocols (González et al, 2002). Serial sections of each brain tissue sample were immunolabelled with four PrP antibodies, namely P4 (mouse monoclonal raised against the 89-104 amino acid sequence of ovine PrP; Hardt et al, 2000), 521.7 and 505.2 (rabbit polyclonals raised against the 94-105 and 100-111 amino acid sequence of ovine PrP, respectively; van Keulen et al, 1996) and R486 (rabbit polyclonal raised against the 221-234 amino acid sequence of bovine PrP; R. Jackman, unpublished).…”

Detection of PrPsc on Lymphoid Tissues from Naturally Affected Scrapie Animals: Comparison of Three Visualization Systems

“…To ensure the specificity of the pathological PrP immunostaining, several pretreatments as previously described (Hardt et al 2000) were performed. Immersion in 98% formic acid for 15 min, proteinase K treatment for 15 min at 37C (Roche, Mannheim, Germany; 4 g/ml), and hydrated autoclaving (immersion in distilled water in a pressure cooker) were applied to all sections before use of any of the visualization systems tested in the present study.…”

“…In this study, different visualization systems used routinely in research and diagnosis (Hardt et al 2000;Lezmi et al 2001;Monleón et al 2003) were assessed to determine whether the methodology applied for IHC examination using lymphoid tissue may alter the results obtained. Furthermore, the study intended to determine which of the systems shows the highest sensitivity, with the aim of providing an accurate tool for investigation of the early pathogenesis of scrapie and, consequently, for preclinical diagnosis.…”

Scrapie in Swine: a Diagnostic Challenge

“…In all hematoxylin and eosin stained slides examined by light microscopy, there was no evidence of spongiform encephalopathy beyond that present in negative control pigs. Examination of slides stained by an immunohistochemical method with monoclonal antibody L42 that targets to amino acids 145-163 of the ovine prion protein sequence 30) demonstrated immunoreactivity that was largely confined within neurons (Fig. 1) and was present in hippocampus, cerebellum, thalamus, brainstem, and spinal cord.…”