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The flow-through containing the target protein was concentrated using AmiconUltra-15 3 K MWCO (Merck KGaA, Darmstadt, Germany) and the concentration was determined from the E280 values determined in a Jasco V-630 Spectrophotometer (Jasco, Grossumstadt, Germany) at 280 nm using the calculated extinction coefficient

Furthermore, iron modulates APP expression and processingenhancing BACE1 activity and thereby increasing A142 production (296)

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For GST fusion proteins, cells were lysed by sonication in Buffer A (50 mM Tris-HCl, pH 8.0, 300 mM NaCl, 1 mM EDTA, 0.5% Triton X-100, 1 mM DTT, 1 mM phenylmethylsulfonyl fluoride/PMSF))