We sampled honey bee queens from beekeepers throughout British Columbia which were rated as either 'failed' or 'healthy' by beekeepers. We performed shot-gun proteomics on the spermathecal fluid (sperm removed). Proteins were reduced, alkylated, and digested with Lys-C for 4 h in urea digestion buffer. The sample was then diluted 4x with ammonium bicarbonated and digested overnight with trypsin. 2 ug of peptides were analyzed by an EASY-nLC 1000 liquid chromatography system coupled to a Bruker Impact II mass spectrometer. Data were searched with MaxQuant and the zipped txt folder output is provided.