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In the next step, samples were diluted with 100 mM ABC, then reduced with 5 mM TCEP (in 50 mM ABC) for 30 min at 37 C, re-alkylated with 10 mM d5-NEM, diluted with four volumes of 25 mM ABC and digested overnight with trypsin at 37 C, (shaking at 550 rpm

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