Liquid chromatography
Separation of peptides was performed as described in Chapter 2.3.4. The LC run started with 10% B for 3 min, followed by a gradient to 30% B in 19 min, another linear gradient step to 100% B in 2 min. An isocratic step at 100% B continued for 7 min. At the end of the run the column was allowed to equilibrate at 10% B for 7 min. The flow rate was 250 μL/min.