Today, the ultimate confirmation of the disease comes only after post-mortem examination of the brain, even though extensive research being carried out in the field offers hope for pre-clinical diagnosis through the detection of PrPC in tonsils or body fluids.
For detection of PrPC and PrPSc, the immunohistochemical procedure was identical but the pretreatment steps differed.
For detection of Prpc and anp32a, brain and spleen tissues were lysed in RIPA buffer (50 mM Tris HCl pH 8, 150 mM NaCl, 1% NP-40, and 0.5% sodium deoxycholate, 0.1% SDS) with protease inhibitors (Sigma P8340; Sigma, St . Louis MO) and centrifuged at 12000 rpm for 20 minutes.
Absence of α-cleavage is characteristic shared by PrPSc [7], [43] and toxic PrPC mutants [14], [15], [16], suggesting that α-cleavage is important for the function of PrPC and possibly even for the pathogenesis of prion diseases.
The detection of native PrPC after enrichment from 10 ml milk from cow, sheep, goat, and human using Alicon PrioTrap® is shown in figure 1.
For detection of PrPC, Cav-1 and CHC, lysates were not treated with PK but were supplemented with 0.5 mM Pefabloc and precipitated with methanol.
Direct interaction of PrPC with gangliosides as well as with p59Fyn kinase in plasma-derived MVs was assayed by coimmunoprecipitation followed either by TLC immunostaining for the detection of gangliosides or conventional immunoblotting for p59Fyn kinase detection.
The different PrPc content in the three tissues of the CNS would be in accordance with the spatiotemporal appearance of PrPSc aggregates and would also support the idea that the obex is the best source of material for the detection of PrPSc in TSE rapid tests analyses.
The increase in susceptibility with age correlated with the immunocytochemical detection of PrPC on maturing FDCs [38].
However, little is known about the physiological role of PrPc and basic PrPc regulating mechanisms.
