The immortalized human neuroblastoma cell line SH-SY5Y has long been utilized for in vitro studies, its usefulness being attributed to its dopaminergic characteristics and ability to differentiate into functional neuron-like cells. Notably, retinoic acid (RA) treatment of SH-SY5Y cells has yielded multiple insights into neuronal differentiation in both the developing and adult brains. In addition, RA-differentiated SH-SY5Y cells provide an in vitro model to study diseases related to abnormalities in dopamine signalling such as Parkinson’s and schizophrenia. Although the proteome of undifferentiated SH-SY5Y cells has been extensively studied, a comprehensive proteomic profile of these cells during and after differentiation is required. Therefore, this study aims to expand on existing proteomic data of RA-differentiated SH-SY5Y cells via quantitative LC-MS/MS. The data generated will enhance understanding of the differentiation process, as well as uncover potential targets for therapeutic agents, with the long-term outcome being improved treatments for relevant neuronal diseases.