In this study, we synthesized copper hydroxide nanosheet and investigated electrochemical oxidation of glucose using the copper hydroxide nanosheets. The precursor of the nanosheet was a layered copper hydroxide synthesized by the ion exchange of dodecylbenzene sulfonate with acetate in Cu2(OH)3(CH3COO)·H2O. Delamination of the layered copper hydroxide prepared the nanosheet by dispersion in 1-butanol. Atomic force microscopy images of the nanosheets showed lateral dimensions of ca. 2 μm with a height of ca. 4.5 nm. Cyclic voltammogram of the nanosheet coated electrode showed oxidation current peak depend on the concentration of glucose at around +0.6 V vs. Ag/AgCl. Amperometry was measured at +0.6V vs. Ag/AgCl with successive addition of glucose solution. Glucose concentration and catalytic current were almost proportional. When the linear range is 0.1 to 4.9 mM, the sensitivity was 1.16 mA mM-1cm-2 from the slope