Creutzfeldt‐Jakob Disease: Carnoy's Fixative Improves the Immunohistochemistry of the Proteinase K‐ Resistant Prion Protein

• DOI: 10.1111/j.1750-3639.2000.tb00240.x
• Publication Date: 2010-07-27T05:35:38Z
• Authors: Giorgio Giaccone; Barbara Canciani; Gianfranco Puoti; Giacomina Rossi; Donato Goffredo; Selina Iussich; Paolo Fociani; Fabrizio Tagliavini; Orso Bugiani
• Keywords: Fixative; Proteinase K; Immunostaining; PRNP; Immunolabeling; Antigenicity

The neuropathological diagnosis of Creutzfeldt‐Jakob disease relies on the immunohistochemical demonstration proteinase‐K resistant form prion protein (PrP res ) in brain tissue. antigenicity PrP is strongly reduced by formalin solution widely used to fix tissue, thus immunoreactivity inconsistently detectable formalin‐fixed A better immunostaining can be obtained using Carnoy's fixing solution, which composed ethanol, chloroform and acetic acid (6:3:1). easily extracted from Carnoy's‐fixed, paraplast‐embedded Accordingly, Carnoy's‐fixed tissue prior immunolabeling subjected proteinase K guanidine thio‐cyanate, respectively eliminate normal cellular promote denaturation. In comparison with best protocols for (i.e. ‐ hydrolytic autoclaving or distilled water followed formic thiocyanate), carried out sections cut blocks thiocyanate, proved more successful detect map both diffuse focal immunoreactivity, correlate pattern MV polymorphism at PRNP codon 129 banding glycosyla‐tion revealed Western blot.