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Hamster brain homogenates were processed by homogenization in a MES buffer (25 mM MES, 150 mM NaCl, 60 mM n-octyl-glucoside and 1% Triton X-100, pH 6.5) at 4 C and protein concentration quantified by BCA (Pierce, IL).
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The resulting homogenates were processed as above.
A volume of 30 μl of cell extracts (approximately 3 × 105 cells) or brain homogenates were intracerebrally inoculated into C57BL/6 mice (group size: 15).
Brain homogenates were assayed according to the manufacturer's instructions.
Mouse brain homogenates were fractionated as described [19].
Virus-containing brain homogenates were obtained as follows.
Normal and Scrapie-infected brain homogenates were incubated with RADA bound to 96-well plates.
To avoid contamination, brain homogenates were prepared in a laboratory that had never contained infected materials.
Brain homogenates were added to 2 ml of β-scintillation liquid (Roth).
Aβ40 and Aβ42 levels in brain homogenates were determined by ELISA.
Mouse brain homogenates were prepared in PBS, 0.05% sodium deoxycholate, and 0.05% Nonidet P-40.
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