Hydrated bentonite is considered an alternative subsurface sealing/plugging material for deep geo-energy wells. However, the knowledge related to this application and the corresponding properties of bentonite is still lacking. This includes the mechanical properties at the interface of bentonite plugs with the adjacent materials (surrounding rock or casing steel) and the mechanical stability of plugs under downhole in-situ conditions. In this work, we performed experiments investigating the interface shear properties and shear strength of a bentonite plug under various settings for deep geo-energy applications, such as hydrocarbon and geothermal wells. The interface’s shear properties against various adjacent materials and fluid conditions were characterized. The influence of chemical exposure, the salinity of the curing fluid, fluid pH, pressure, and temperature on bentonite’s mechanical stability was evaluated in a small- and large-scale setting. The latter was performed using realistic casing sizes and placement methods, relevant for the field application. The experimental results show that the averaged shear strength of the bentonite plug interface is 13.3 kPa and 9.1 kPa when cured in freshwater and seawater, respectively. The increase in strength with increasing curing pressure, temperature, and fluid pH was characterized for the first time. The interfacial properties of cohesion and friction angle vary with different surrounding materials. They are also influenced by the saturating condition and salinity of the saturating fluid. Based on the experimental results, a bentonite plug with a minimum length of 15–43 m placed in casings of 7–5/8″ to 13–3/8″ would be sufficient to meet the necessary criteria of the Dutch regulators as an isolating material for well abandonment.