Immunoreactivity of specific epitopes of PrPSc is enhanced by pretreatment in a hydrated autoclave.
An abnormal protein (PrPSc) accumulates in animals affected with scrapie. Immunoblotting procedures have been used widely to detect PrPSc. Blotted membranes were subjected to pretreatment in a hydrated autoclave, and the subsequent immunoreactivity of PrPSc was examined. The immunoreactivity of PrPSc to antisera against the synthetic peptides of the mouse PrP amino acid sequences 199 to 208 and 213 to 226 was enhanced by the pretreatment. However, the reactivity to antisera of peptide sequences 100 to 115 and 165 to 174 was not affected. The antibody-binding ability of the specific epitopes which are located close to the C-terminal end of PrP27-30 the proteinase-resistant portion of PrPSc, was enhanced by pretreatment in a hydrated autoclave. This pretreatment increased the sensitivity of PrPSc, and it would be useful for diagnosis of scrapie.
Yokoyama
T
T
National Institute of Animal Health, Ibaraki, Japan. tyoko@niah affrc.go.jp
Momotani
E
E
Kimura
K
K
Yuasa
N
N
D000818:Animals; D000939:Epitopes / Q000032:analysis* / Q000276:immunology / Q000528:radiation effects*; D006358:Hot Temperature; D015151:Immunoblotting / Q000379:methods; D051379:Mice; D018620:PrPSc Proteins / Q000276:immunology* / Q000528:radiation effects*; D012608:Scrapie / Q000276:immunology; D013242:Sterilization / Q000295:instrumentation*; D014867:Water / Q000737:chemistry*
D000939:Epitopes; D018620:PrPSc Proteins; D014867:Water