We describe the first comprehensive venom profile of a solitary bee, the European carpenter bee Xylocopa violacea, by using a combined proteo-transcriptomic approach. Transcripts of the venom system are used as specific database to match peptide sequences from mass spectrometry analyses of venom from dissected venom glands and venom injected during a defensive sting. In our analyses, we only discuss the proteo-transcriptomically supported venom components and reveal a diverse venom profile with 43 different protein families of which 32 are identified in the injected venom.