Abstract
Western immunoblotting is a workhorse technique used in the prion field to analyze disease-associated forms of the prion protein, termed PrPSc. The biochemical stability of PrPSc aggregates combined with the increased resistance of prion infectivity to inactivation by various treatments that inactivate most other pathogens complicates the use of Western immunoblotting as a means to characterize PrPSc samples. In this chapter, we describe a method for Western immunoblot analysis of PrPSc with an emphasis on precautions to address the biochemical and biosafety considerations associated with this procedure.
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Acknowledgment
This research was supported by the Intramural Research Program of the National Institute of Allergy and Infectious Diseases, National Institutes of Health. Animal experiments were conducted in an Association for Assessment and Accreditation of Laboratory Animal Care International (AAALAC)-accredited facility in accordance with animal welfare guidelines under an animal study protocol (2009-27) approved by the Animal Care and Use Committee of the Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health.
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Baron, G.S., Raymond, G.J. (2017). Immunodetection of PrPSc Using Western Immunoblotting Techniques. In: Lawson, V. (eds) Prions. Methods in Molecular Biology, vol 1658. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7244-9_5
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DOI: https://doi.org/10.1007/978-1-4939-7244-9_5
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