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Protein pellets were suspended and denatured in 20 l 7 M urea/2 M thiourea/100 mM TEAB, pH 7.5, reduced with 2 L of 50 mM Tris(2-carboxyethyl) phosphine (TCEP, SCIEX), pH 8.0, at 37 C for 60 min and followed by the addition of 1 L of 200 mM cysteine-blocking reagent (methyl methanethiosulfonate (MMTS, Pierce) for 10 min at room temperature

Blood was obtained via cardiac puncture and centrifuged at 4000 rpm at 4 C for 10 min to collect plasma

In all that time harmful effects have never been shown from overdose

Changes in the makeup or function of gut bacteria could compromise the intestinal barrier, allowing prions to travel from the gut to the brain