The flip-chip technology using anisotropic conductive films (ACFs) is gaining growing interest due to its technical advantages such as environmentally friendly, simpler, and lower cost processes. Electrical performances and reliability of ACF flip-chip assembly depend on thermomechanical properties of ACF polymer resins. In this paper, the changes in ACF resin morphology due to the phase separation of thermoplastics, and subsequent changes of physical and mechanical properties were investigated as a function of thermoplastic contents of ACF formulation. Furthermore, the pressure cooker test (PCT) reliability of ACF flip-chip assemblies with various thermoplastic contents was also investigated. As thermoplastic contents increased, coefficient of thermal expansion (CTE) of ACFs increased, and elastic modulus (E') of ACFs decreased. In contrast, water absorption rate decreased as thermoplastic content increased. As a result, PCT reliability of ACF flip-chip assembly was improved adding up to 50 wt.% content of thermoplastic.