Regular ArticleExpression of Human Amyloid Precursor Protein Ectodomains inPichia pastoris:Analysis of Culture Conditions, Purification, and Characterization☆
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Evaluation of the effects of treatment with sAPPα on functional and histological outcome following controlled cortical impact injury in mice
2012, Neuroscience LettersCitation Excerpt :This time point has been previously used to examine the effects of sAPPα treatment on functional and histological outcome following a diffuse injury in rats [6,31], with ICV administration of sAPPα shown to result in a homogenous distribution of sAPPα throughout the rat brain and at a higher concentration compared to shams [27]. The sAPPα was produced as previously described, expressed as a secreted protein in the methylotrophic yeast, Pichia pastoris [15]. 150 μL of 25 μM sAPPα was then combined with 75 μL of aCSF to produce the final solution used to treat the mice.
α-secretase-derived fragment of cellular prion, N1, protects against monomeric and oligomeric amyloid β(Aβ)-associated cell death
2012, Journal of Biological ChemistryCitation Excerpt :Protein expression was induced during 48 h in BMMY (1% yeast extract, 2% peptone, 1.34% yeast nitrogen base without amino acids (Sigma), 4 × 10−5% biotin (Sigma), 2% methanol (Merck)). Purification was carried out on ice using a modification of the method of Henry et al. (30). Yeast cultures (0.5–2 liters) were centrifuged at 16,000 × g for 10 min at 4 °C, and supernatants were filtered (0.45 μm; Whatman).
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J. E. Coligan, Ed.
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