Detection of glucose is highly informative, creating a constant demand for fabricating high-precision glucose biosensors. Metal–organic frameworks, a family of porous materials renowned for their tunability, can be an excellent choice for developing such sensors. We have developed a highly-sensitive, non-enzymatic sensor for electrochemical detection of glucose fabricated using Copper Metal–Organic Framework (Cu MOF), synthesized by a simple, room-temperature stirring method using Benzene-1,3,5-tricarboxylic acid (BTC) as ligand and Copper nitrate trihydrate as precursor. The synthesized nanostructure was characterized using Fourier transform infrared spectroscopy, scanning electron microscopy, and energy-dispersive X-ray analytical techniques. Powder X-ray diffraction study and thermogravimetric analysis were also done. Further, Brunnauer-Emmett-Teller analysis revealed the porous nature of Cu MOF. The materials exhibited strong electro-catalytic activity for glucose oxidation as revealed from cyclic voltammetry and chronoamperometric studies done under alkaline pH conditions. The Cu MOF deposited on a conducting graphite sheet electrode displayed a significantly low detection limit of 0.019 mM through a broad detection range (1–15 mM) and a strong sensitivity of 229.4 μAmM−1 cm2. Overall, the Cu MOF/GS exhibits exceptional stability, short response time (less than 1 s), and good repeatability and reproducibility, making it a promising future material for non-enzymatic glucose detection.