Reproducible, simplified deep proteomic coverage remains one of the significant challenges and desires for non-specialist MS researchers. Although many different approaches are available for these purposes, few are applicable in day-to-day, standardized, and easy-to-handle fashion. Most methods require specialist knowledge and development skills to achieve suitable results. Standard methods for using high pH fractionation are on reversed-phase sorbents with buffers containing variable acetonitrile percentages potentially introducing sources of error. Here, we present a novel cartridge-based fractionation using an alternative chemical separation and dedicated buffer compositions. The fractionation gave up to 1.7-fold increase in protein and peptide identifications for S. cerevisiae samples. Additional experiments on blood plasma and HeLa cell samples were performed, demonstrating a similar increase in protein identification rate compared to traditional iST workflow. Overall, the method fractionation only adds 10 min to traditional iST workflow with a substantial increase in protein identifications for three different matrices. The fractionation add-on to the iST kit is easy-to-use, suitable for automation platforms and high-throughput workflows.