Urea-containing buffer solutions are generally used in proteomic studies to aid protein denaturation and solubilisation during cell and tissue lysis. It is well-known, however, that urea can lead to the carbamylation of peptides and proteins and; subsequently, incomplete digestion of proteins. In our work, we sought for ways to retain urea in the digestion protocol to reduce sample handling, while minimizing urea-related drawbacks.