Immunodetection of PrPSc using Western and Slot blotting techniques.
Prion infectivity is often linked to presence of the protease-resistant isoform of prion protein (PrP), PrP(res); therefore, it is of highest interest to have convenient methods for rapid detection of PrP(res) in the research laboratory. For detection of PrP(res) in model systems to confirm infectivity, there are several methods that can be applied. This chapter focuses on detection of PrP(res) by proteinase K digestion followed by Western blot, which is the only method that is both quantitative and qualitative. For large-scale screening of PrP(res) content in samples, the dot blot method offers a great advantage for detecting PrP(res), and this method is also thoroughly described in this chapter.
Gyllberg
Hanna
H
Department of Biochemistry and Biophysics, Stockholm University, Sweden.
Löfgren
Kajsa
K
D015153:Blotting, Western / Q000379:methods*; D019286:Endopeptidase K / Q000378:metabolism; D047468:Immunoprecipitation; D008567:Membranes, Artificial; D018620:PrPSc Proteins / Q000032:analysis*
D008567:Membranes, Artificial; D018620:PrPSc Proteins; D019286:Endopeptidase K