Developing facile strategies to construct free-standing flexible electrodes with extremely sensitive and stable electrochemical response towards analytes is of huge demand and it is still highly challenging. Herein, we design a unique and cost-effective hybrid electrode comprising of cobalt oxide (Co3O4) directly grown on highly conducting MXene based carbon fiber electrode (CFE) as a self-standing electrode using a simple hydrothermal-annealing method, where MXene nanosheets on electrode support facilitate the growth of uniform-sized Co3O4 nanocubes. Our results demonstrate that the synergetic effects of conductive MXene support and catalytically active Co3O4 nanocubes provide a superior response towards glucose oxidation in an alkaline medium (0.1 M KOH). The resultant Co3O4/[email protected] demonstrates remarkable sensitivity (19.3 μA mM−1 cm−2), wide linear range (0.05 μM to 7.44 mM) and lower detection limits (10 nM), thus providing excellent selectivity over interfering species and good electrochemical stability for an extended shelf-life. Therefore, the obtained self-standing Co3O4/[email protected] flexible electrode establishes a promising platform for glucose oxidation and it could be envisioned for the highly sensitive detection of various target molecules for their widespread applications in clinical diagnosis and health care monitoring.