Updated project metadata. Protein termini play pivotal roles in various biological processes. Although a series of terminomic strategies have been proposed for the analysis of protein N-termini or protein C-termini separately, few can analyze both ends of proteins at the same time. Herein, we developed a workflow, termed Simultaneous Analysis of Protein N- and C- Terminome (SAPT) based on strong cation exchange chromatography (SCX) to study protein terminome simultaneously.