Ceramide is a bioactive molecule involved in numerous cell signaling pathways that are associated with cell cycle control, differentiation, senescence, and apoptosis. Although substantial knowledge about ceramide‐regulated pathways has accumulated in the past decade, molecular mechanisms of ceramide action remain poorly understood, primarily due to limited information about ceramide‐binding proteins. In the present study, we used affinity purification with a synthetic biotin‐conjugated C6‐ceramide analogue and LC‐MS/MS to identify potential ceramide‐interacting proteins in D6P2T Schwannoma cells. The purification resulted in identification of 97 unique proteins. The identified proteins are involved in various cellular processes, including apoptosis, cellular stress, cell cycle, cell differentiation, signaling, transcription, translation, protein biogenesis, metabolism, and transport.