The medial collateral ligament is one of the most frequently injured ligaments in the knee. Although the medial collateral ligament is known to provide a primary restraint to valgus and external rotations, details regarding its precise mechanical function are unknown. In this study, strain in the medial collateral ligament of eight knees from male cadavers was measured during valgus loading. A material testing machine was used to apply 10 cycles of varus and valgus rotation to limits of ±10.0 N-m at flexion angles of 0°, 30°, 60°, and 90°. A three-dimensional motion analysis system measured local tissue strain on the medial collateral ligament surface within 12 regions encompassing nearly the entire medial collateral ligament surface. Results indicated that strain is significantly different in different regions over the surface of the medial collateral ligament and that this distribution of strain changes with flexion angle and with the application of a valgus torque. Strain in the posterior and central portions of the medial collateral ligament generally decreased with increasing flexion angle, whereas strain in the anterior fibers remained relatively constant with changes in flexion angle. The highest strains in the medial collateral ligament were found at full extension on the posterior side of the medial collateral ligament near the femoral insertion. These data support clinical findings that suggest the femoral insertion is the most common location for medial collateral ligament injuries.