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After reduction (10 mM dithiothreitol, 30 min at room temperature), alkylation (20 mM iodoacetamide, 30 min at room temperature in the dark), and quenching of excess iodoacetamide with 5 mM dithiothreitol (10 min at room temperature in the dark), the sample was diluted into 4 M urea with 50 mM ABC before adding 1 g of endoproteinase Lys-C (Wako) for protein digestion

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