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This denaturation is often done deliberately in the course of separating and purifying proteins
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After 2 days of expression, cells were harvested in ice-cold PBS and subsequently lysed in AP-MS buffer (10 mM Tris/Cl pH 7.5, 150 mM NaCl, 10% Glycerol, 0.5 mM EDTA, 0.5% Nonidet P40) supplemented with protease inhibitor cocktail (Roche, Rotkreuz, Switzerland) by sonification, followed by centrifugation (20,000 g for 10 min
If even one amino acid is out of place, or if the sample is contaminated with solvents, salts, or incomplete fragments from the synthesis process, the results are compromised