Updated FTP location. In this project, we developed a rapid enzymatic digestion protocol within an anion exchange StageTip named AnExSP (anion-exchange disc-assisted sequential sample preparation) and, using data-independent acquisition, analyzed its digestion performance of human cellular protein extracts (HCPE) using trypsin and trypsin/Lys-C. The outstanding effectiveness and simplicity of AnExSP in sample preparation for gel-based cross-linking mass spectrometry was also verified.