title: Effective antigen-retrieval method for immunohistochemical detection of abnormal isoform of prion proteins in animals.
abstract: For immunohistochemistry of the prion diseases, several pretreatment methods to enhance the immunoreactivity of human and animal abnormal proteinase-resistant prion protein (PrP(Sc)) on the tissue sections have been employed. The method of 121 degree C hydrated autoclaving pretreatment or the combination method of 121 degree C hydrated autoclaving with a certain chemical reagent (formic acid or proteinase K, etc) are now widely used. We found that an improved hydrated autoclaving method at 135 degrees C, more effectively enhanced PrP(Sc) immunoreactivity for the antibodies recognizing the linear epitope. In addition, this method was more effective for the long-term fixation samples as compared with other previous methods. However, this modified method could not retrieve PrP(Sc) antigenic epitopes composed of conformational structures or several discontinuous epitopes. We describe the comparative studies between our improved method and other antigen-retrieval procedures reported previously. Based on the differences of reaction among the antibodies, we also discuss the mechanisms of the hydrated autoclaving methods to retrieve PrP(Sc) immunoreactivity.
authors.affiliation: Department of Pathobiological Science, Obihiro University of Agriculture and Veterinary Medicine, 080-8555, Obihiro, Japan. furuoka@obihiro.ac.jp
authors.forename: Hidefumi
authors.initials: H
authors.lastname: Furuoka
authors.forename: Atushi
authors.initials: A
authors.lastname: Yabuzoe
authors.forename: Motohiro
authors.initials: M
authors.lastname: Horiuchi
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Tagawa
authors.forename: Takashi
authors.initials: T
authors.lastname: Yokoyama
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Yamakawa
authors.forename: Morikazu
authors.initials: M
authors.lastname: Shinagawa
authors.forename: Tetsutaro
authors.initials: T
authors.lastname: Sata
mesh_terms: D000818:Animals; D000937:Antigen-Antibody Reactions; D015153:Blotting, Western / Q000379:methods; D001921:Brain / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D002417:Cattle; D000939:Epitopes / Q000378:metabolism; D016591:Histocytological Preparation Techniques / Q000662:veterinary*; D007150:Immunohistochemistry / Q000379:methods / Q000662:veterinary*; D051379:Mice; D008813:Mice, Inbred ICR; D011328:Prions / Q000032:analysis* / Q000276:immunology; D020033:Protein Isoforms / Q000032:analysis* / Q000276:immunology; D012756:Sheep; D013045:Species Specificity; D013696:Temperature
chemical_list: D000939:Epitopes; D011328:Prions; D020033:Protein Isoforms