Amphiphilic nanosheets have gained great attraction in diverse applications owing to their unprecedented performance. In this paper, amphiphilic molybdenum disulfide (CTAB−MoS 2 ) nanosheets were prepared through a one-step simple hydrothermal method, and the physicochemical properties were systematically investigated. The results showed that the CTAB−MoS 2 nanosheets displayed a distinct ultrathin flakelike structure and a lateral size in the range of 95 nm. More specifically, the presence of defect sites over the surfaces of MoS 2 nanosheets facilitated linkage with CTAB molecules, which made MoS 2 amphiphilic. Because of the amphiphilic nature, the CTAB− MoS 2 nanosheets could be steadily dispersed in water phase, reduce the oil−water interfacial tension, alter wettability of the solid surface, and stabilize emulsions at ultralow concentrations (50 mg/L). By observing and analyzing the interfacial behavior of CTAB−MoS 2 nanosheets in a simulated oil/brine system, it was found that CTAB−MoS 2 nanosheets could be adsorbed at the oil− water interface in the form of multilayer adsorption, and the adsorption layer (oil−water interfacial film) exhibited higher strength elasticity. This work provides a scalable method for preparing amphiphilic nanosheets and reveals the physicochemical properties of CTAB−MoS 2 amphiphilic nanosheets.