Layered silicate AMH-3 forms a 3D ordered microporous structure and is potentially useful as a permselective barrier. Nafion-based composite membrane containing delaminated AMH-3 (D-AMH-3) layer was prepared by solution casting and hot pressing. The membrane structure was analyzed by FE-SEM and EDS, revealing a sandwich-type structure that included double Nafion outer layers and a central D-AMH-3 layer. The Nafion/D-AMH-3 membrane was employed as an ion exchange membrane for VRB application, and the vanadium permeability and single cell performance were evaluated. The Nafion/D-AMH-3 membrane exhibited a lower VO2+ permeability compared to N117, resulting in higher Coulombic efficiency and lower capacity loss per cycle. The results indicated that D-AMH-3 layer is potentially suitable as a permselective barrier for reducing vanadium crossover and improving cell performance. •Nafion/D-AMH-3 membrane had a sandwich structure.•D-AMH-3 layer acted as a permselective barrier to reduce vanadium crossover.•Nafion/D-AMH-3 membrane improved the Coulombic efficiency and capacity retention.