In this study, we developed a novel free-standing sensor based on hierarchical zinc-doped nickel hydroxide nanosheets vertically attached three-dimensional (3D) framework (Zn-doped Ni(OH)2 NSs/3DF) via a mild and facile hydrothermal reaction for non-enzymatic glucose detection. The Zn-doped Ni(OH)2 NSs/3DF-based sensor exhibited a good sensitivity of 52μAμM−1 cm−2, wide linear detection range of 7.9nM to 15.2μM, and a low detection limit of 26.6nM. Besides, the respectable selectivity and excellent stability of such sensor were also demonstrated. The achieved results were due to the formation of the Zn-doped Ni(OH)2 NSs, which played an important role for promoting the electroactive sites and surface area, leading to the improvement of electrocatalytic activity towards glucose oxidation. In addition, the successful fabrication of a unique self-supported 3D hierarchical architecture not only produced the enhanced charge conductivity but also assisted to the sensing stability because it effectively avoided the agglomeration or exfoliation of NSs. The remarkable sensing activity and stability of the Zn-doped Ni(OH)2 NSs/3DF-based sensor offered a facile and appropriate approach towards glucose detection, which is highly promising to apply for practical clinical analyses.