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For trypsin digestion, the beads were resuspended in 200 L of 2 M urea in Ca- and Mg-free D-PBS, 2 L of 100 mM calcium chloride (CaCl, Sigma-Aldrich, UK) to achieve a final concentration of 1 mM, and 4 L of 0.5 mg/mL trypsin (2 g total