2.2. Trypsin Digestion

The protein sample was incubated with dithiothreitol (5 mM) at 56 °C for 30 min. Then, the protein sample was supplemented with iodoacetamide (11 mM) in the dark at room temperature for 15 min and lysed. Later, the samples were added with 100 mM TEAB, keeping the urea concentration below 2 M. Finally, trypsin was used for final digestion at two different mass ratios. First, digestion was performed at a 1:50 ratio overnight, followed by a 1:100 mass ratio for 4 h.