Objective To establish a stable PrPSc panel from brain tissues of experimental hamsters infected with scrapie agent 263K for evaluating diagnostic techniques of human and animals' prion diseases. Methods Thirty brain tissue samples from hamsters intracerebrally infected with scrapie strain 263K and another 30 samples from normal hamsters were selected to prepare 10%, 1%, and 0.5% brain homogenates, which were aliquoted into stocks. PrPSc in each brain homogenate was determined by proteinase K digestions followed by Western blot assay and partially by immunohistochemistry. Stability and glycoforms of PrPSc were repeatedly detected by PrPSc-specific Western blots in half a year and 3 years later. Results PrPSc signals were observed in all 10% brain homogenates of infected hamsters. Twenty out of 30 stocks and 19 out of 30 stocks were PrPSc positive in 1% and 0.5% brain homogenatesof infected hamsters, respectively. Twenty-seven out of 30 stocks presented three positive bands in 10% brain homogenates, whereas none of 1% and 0.5% homogenates contained 3 bands. The detection of PrPSc-specific signals stored in half a year and 3 years later demonstrated that the ratio of PrPSc positive samples and glycoforms was almost unchanged. All normal hamsters' brain homogenates were PrPSc negative. Conclusion A PrPSc panel of prion disease can be established, which displays reliably stable PrPSc-specific signals and glycoforms.