In this study, an ultrasensitive label‐free electrochemical immunosensor based on Ti3C2Tx (Mxene) with a 2D‐layered morphology has been proposed for the detection of EpCAM antigen. A hydrothermal method is employed for the synthesis of Ti3C2Tx by using a less toxic exfoliating reagent, NaBF4. The bioelectrode, BSA/anti‐EpCAM/Ti3C2Tx@ITO, has been fabricated by electrophoretic deposition of Ti3C2Tx onto the ITO electrode, followed by an immobilization of EpCAM antibody. Electrochemical response studies reveal that the immunosensor shows a high sensitivity of 29.22 μA fg−1 ml cm−2 and a wide linear range from 0.1 fg/ml to 100 ng/ml, for EpCAM antigen detection. Further, the good stability of this fabricated immunosensor is an additional advantage for EpCAM antigen detection in serum samples.