Proteins are essential regulators of biological processes and can have diverse functions depending on their interactions with other proteins. To understand deeply the interaction and their complexes, co-fractionation mass spectrometry (CF-MS) was used with ion exchange chromatography columns and data was analyzed with Data-independent acquisition (DIA) mode. Our comprehensive DIA CF-MS analysis successfully identified over 1,000 protein complexes and approximately 10,000 protein-protein interactions, many of which had not been previously discovered. These findings present an invaluable resource for a wide range of human brain research endeavors, including the identification of disease-specific protein complexes.