Bile is an important body fluid with crucial functions ranging from fat absorption to excretion of metabolic breakdown products. Although the chemical composition of human bile is well understood, its protein constituents are beginning to be unravelled only recently. Here, we provide an overview of the proteomics of human bile, both in health and in diseases such as cholangiocarcinoma. A comprehensive catalog of proteins in the bile should facilitate better understanding of its role in physiology and the development and validation of biomarkers for hepatobiliary disorders.