Prion Protein Analysis Techniques

Three dyes, ThT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and DAPI, and two antibodies, the mouse anti-PrP monoclonal antibody 3F4^{41 (link),48 (link)} and a 10-nm gold-labeled anti-mouse antibody^{7 ,9 (link),57 (link)}, were purchased from Sigma-Aldrich (St. Louis, MO). Sarkosyl and PK were obtained from Amresco (Solon, OH). Alexa 546-conjugated fluorescent secondary antibody^{7 ,9 (link),57 (link)}, the proteasome inhibitor MG132, and the ROS probe DCFH-DA (2′,7′-dichlorofluorescein diacetate) were purchased from Beyotime (Nantong, China), respectively. Ni-Sepharose was obtained from GE Company (Pittsburgh, PA). All other chemicals used in this study were of analytical grade and were produced in China.