The identification of biological stains and their tissue resource is an important part of forensic research. Current methods suffer from several limitations including poor sensitivity and specificity, trace samples, and sample destruction. In this study, we profiled the proteomes of menstrual blood, peripheral blood, saliva, semen, and vaginal fluid with mass spectrometry technology. Tissue-enhanced and tissue-specific proteins of each group have been proposed as potential biomarkers. These candidate proteins were further annotated and screened through the combination with the human protein atlas database. Our data not only validates the protein biomarkers reported in previous studies but also identifies new candidate biomarkers for human body fluids. These candidate protein biomarkers lay the foundation for the development of rapid and specific forensic examination methods.