Mice are commonly considered the model mammal for many biomedical studies. In this work, mouse and human dentin were compared to specify structural and mechanical differences to establish a baseline for comparison of dental tissues between these species. Atomic force microscopy revealed tubules of about 1.0 to 1.6 μm in diameter as the main structural feature in dentin of both species. Nanoindentation yielded the elastic modulus about 15% lower in murine intertubular dentin while the hardness was almost equal. Dynamic stiffness mapping confirmed the lower elastic properties and also revealed that the peritubular region of increased mineralization around tubules is drastically reduced or maybe absent in murine dentin of this age. This study suggests that structural and mechanical differences need to be considered when murine dentin is used as a model system.