Samples were obtained from HuBMAP Tissue Mapping Centers from ten human donors. Tissue was cryopulverized, lysed, and proteins precipitated (Figure ). To increase the depth of proteome coverage, proteins were fractionated with GELFrEE prior to MS analysis. Since we intended to analyze each sample by both CZE and RPLC, we setup two Orbitrap tribrid MS instruments configured with either CZE or RPLC, acquired data for a sample on one system, and immediately acquired data for the same sample on the second one. CZE substantially benefits from a higher scan rate due to generally narrower peak widths. Consequently, the CESI 8000 Plus was hyphenated to the Orbitrap Eclipse while a Dionex nanoLC was coupled to the Orbitrap Fusion Lumos. Three tissue types (heart, small intestine, and spleen) were analyzed by this paired analysis while two tissues (lung and kidney) were analyzed solely by RPLC-MS on the Orbitrap Eclipse (Table ).