The data presented here is related to the research article entitled "Characterization of the gila monster (Heloderma suspectum suspectum) venom proteome" by Sanggaard et al. in Journal of Proteomics [1]. The gila monster venom was collected, analyzed by 2D-gel electrophoresis and after Coomassie-Brilliant Blue staining the major spots were excised, subjected to in-gel trypsin digestion, and analyzed by LC-MS/MS. Subsequently, the venom proteins were identified based on de novo sequencing and homology searching. The mass spectrometry proteomics data have been deposited to the ProteomeXchange (dataset identifier PXD0001343), and in the present article we present an overview of the identified proteins. Protein identification failed for three of the selected spots, with the method described above. Instead, an iterative process, based on de novo sequencing, was employed.