In this study, the force and stress distribution within the anteromedial (AM) and posterolateral (PL) bundles of the anterior cruciate ligament (ACL) in response to an anterior tibial load with the knee at full extension was calculated using a validated three-dimensional finite element model (FEM) of a human ACL. The interaction between the AM and PL bundles, as well as the contact and friction caused by the ACL wrapping around the bone during knee motion, were included in the model. The AM and PL bundles of the ACL were simulated as incompressible homogeneous and isotropic hyperelastic materials. The multiple-degrees-of-freedom (DOF) knee kinematics of a cadaveric knee were first obtained using a robotic/universal force–moment sensor testing system. These data were used as the boundary conditions for the FEM of the ACL to calculate the forces in the ACL. The calculated forces were compared to the in situ force in the ACL, determined experimentally, to validate the model. The validated FEM was then used to calculate the force and stress distribution within the ACL under an anterior tibial load at full extension. The AM and PL bundles shared the force, and the stress distribution was non-uniform within both bundles with the highest stress localized near the femoral insertion site. The contact and friction caused by the ACL wrapping around the bone during knee motion played the role of transferring the force from the ACL to the bone, and had a direct effect on the force and stress distribution of the ACL. This validated model will enable the analysis of force and stress distribution in the ACL in response to more complex loading conditions and has the potential to help design improved surgical procedures following ACL injuries.