{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2023,4,13]],"date-time":"2023-04-13T22:19:12Z","timestamp":1681424352157},"reference-count":9,"publisher":"Canadian Science Publishing","issue":"6","license":[{"start":{"date-parts":[[2008,6,1]],"date-time":"2008-06-01T00:00:00Z","timestamp":1212278400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/www.nrcresearchpress.com\/page\/about\/CorporateTextAndDataMining"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Can. J. Microbiol."],"published-print":{"date-parts":[[2008,6]]},"abstract":"<jats:p> Detection of the abnormal or the pathogenic form of prion protein (PrP<jats:sup>Sc<\/jats:sup>) by Western blot (WB) is challenging, especially, for samples derived from cell cultures that contain low levels of PrP<jats:sup>Sc<\/jats:sup>. A variety of PrP<jats:sup>Sc<\/jats:sup> concentration methods have been reported with various PrP<jats:sup>Sc<\/jats:sup> recovery efficiencies. Ultracentrifugation is one of the methods used frequently to enrich the pathogenic form of PrP<jats:sup>Sc<\/jats:sup> prior to WB analyses. The resulting PrP<jats:sup>Sc<\/jats:sup> pellet is extremely insoluble and often requires sonication to be dissolved, potentially generating aerosols. We modified the common protein-precipitating protocol using trichloroacetic acid to concentrate PrP<jats:sup>Sc<\/jats:sup> by slow-speed centrifugation, followed by solubilization of the pellets with 6\u00a0mol\/L urea prior to sodium dodecyl sulphate \u2013 polyacrylamide gel electrophoresis and WB analyses. Comparative studies suggest this simple trichloroacetic acid protocol was more effective in enriching PrP<jats:sup>Sc<\/jats:sup> presented in cell cultures and brain homogenates than other reported protein-precipitating methods. Furthermore, incorporation of the urea treatment step to dissolve the precipitated PrP<jats:sup>Sc<\/jats:sup> pellets helped to reduce the infectivity of PrP<jats:sup>Sc<\/jats:sup>. <\/jats:p>","DOI":"10.1139\/w08-027","type":"journal-article","created":{"date-parts":[[2008,6,5]],"date-time":"2008-06-05T18:12:43Z","timestamp":1212689563000},"page":"467-471","source":"Crossref","is-referenced-by-count":5,"title":["Comparison of trichloroacetic acid with other protein-precipitating agents in enriching abnormal prion protein for Western blot analysis"],"prefix":"10.1139","volume":"54","author":[{"given":"Matthew","family":"LeBrun","sequence":"first","affiliation":[{"name":"Center for Biologics Research, Biologics and Genetic Therapies Directorate, Health Canada, 251 Sir Frederick Banting Driveway, A\/L 2201C, Ottawa, ON K1A\u00a00K9, Canada."},{"name":"Ottawa Laboratory\u00a0\u2013 Fallowfield, Canadian Food Inspection Agency, Ottawa, ON K2H 8P9, Canada."},{"name":"National Microbiology Laboratory, Health Canada, Winnipeg, MB R3E 3R2, Canada."},{"name":"Biochemistry, Microbiology and Immunology, University of Ottawa, Ottawa, ON K1H 8M5, Canada."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hongsheng","family":"Huang","sequence":"additional","affiliation":[{"name":"Center for Biologics Research, Biologics and Genetic Therapies Directorate, Health Canada, 251 Sir Frederick Banting Driveway, A\/L 2201C, Ottawa, ON K1A\u00a00K9, Canada."},{"name":"Ottawa Laboratory\u00a0\u2013 Fallowfield, Canadian Food Inspection Agency, Ottawa, ON K2H 8P9, Canada."},{"name":"National Microbiology Laboratory, Health Canada, Winnipeg, MB R3E 3R2, Canada."},{"name":"Biochemistry, Microbiology and Immunology, University of Ottawa, Ottawa, ON K1H 8M5, Canada."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Runtao","family":"He","sequence":"additional","affiliation":[{"name":"Center for Biologics Research, Biologics and Genetic Therapies Directorate, Health Canada, 251 Sir Frederick Banting Driveway, A\/L 2201C, Ottawa, ON K1A\u00a00K9, Canada."},{"name":"Ottawa Laboratory\u00a0\u2013 Fallowfield, Canadian Food Inspection Agency, Ottawa, ON K2H 8P9, Canada."},{"name":"National Microbiology Laboratory, Health Canada, Winnipeg, MB R3E 3R2, Canada."},{"name":"Biochemistry, Microbiology and Immunology, University of Ottawa, Ottawa, ON K1H 8M5, Canada."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stephanie","family":"Booth","sequence":"additional","affiliation":[{"name":"Center for Biologics Research, Biologics and Genetic Therapies Directorate, Health Canada, 251 Sir Frederick Banting Driveway, A\/L 2201C, Ottawa, ON K1A\u00a00K9, Canada."},{"name":"Ottawa Laboratory\u00a0\u2013 Fallowfield, Canadian Food Inspection Agency, Ottawa, ON K2H 8P9, Canada."},{"name":"National Microbiology Laboratory, Health Canada, Winnipeg, MB R3E 3R2, Canada."},{"name":"Biochemistry, Microbiology and Immunology, University of Ottawa, Ottawa, ON K1H 8M5, Canada."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aru","family":"Balachandran","sequence":"additional","affiliation":[{"name":"Center for Biologics Research, Biologics and Genetic Therapies Directorate, Health Canada, 251 Sir Frederick Banting Driveway, A\/L 2201C, Ottawa, ON K1A\u00a00K9, Canada."},{"name":"Ottawa Laboratory\u00a0\u2013 Fallowfield, Canadian Food Inspection Agency, Ottawa, ON K2H 8P9, Canada."},{"name":"National Microbiology Laboratory, Health Canada, Winnipeg, MB R3E 3R2, Canada."},{"name":"Biochemistry, Microbiology and Immunology, University of Ottawa, Ottawa, ON K1H 8M5, Canada."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xuguang","family":"Li","sequence":"additional","affiliation":[{"name":"Center for Biologics Research, Biologics and Genetic Therapies Directorate, Health Canada, 251 Sir Frederick Banting Driveway, A\/L 2201C, Ottawa, ON K1A\u00a00K9, Canada."},{"name":"Ottawa Laboratory\u00a0\u2013 Fallowfield, Canadian Food Inspection Agency, Ottawa, ON K2H 8P9, Canada."},{"name":"National Microbiology Laboratory, Health Canada, Winnipeg, MB R3E 3R2, Canada."},{"name":"Biochemistry, Microbiology and Immunology, University of Ottawa, Ottawa, ON K1H 8M5, Canada."}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"155","reference":[{"key":"atypb1\/ref1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.74.9.4377-4386.2000"},{"key":"atypb2\/ref2","first-page":"193","volume":"69","author":"Huang H.","year":"2005","journal-title":"Can. 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