Representative data of asymptomatic, native-knee kinematics is important when studying changes in knee function across the lifespan. High-speed stereo radiography (HSSR) provides a reliable measure of knee kinematics to <1 mm of translation and 1° of rotation, but studies often have limited statistical power to make comparisons between groups or measure the contribution of individual variability. The purpose of this study is to examine in vivo condylar kinematics to quantify the transverse center-of-rotation, or pivot, location across the flexion range and challenge the medial-pivot paradigm in asymptomatic knee kinematics. We quantified the pivot location during supine leg press, knee extension, standing lunge, and gait for 53 middle-aged and older adults (27 men;​ 26 women:​ 50.8 ± 7.0 yrs, 1.75 ± 0.1 m, 79.1 ± 15.4 kg). A central- to medial-pivot location was identified for all activities with increased knee flexion associated with posterior translation of the center-of-rotation. The association between knee angle and anterior-posterior center-of-rotation location was not as strong as the relation between medial–lateral and anterior-posterior location, excluding gait. The Pearson’s correlation for gait was stronger between knee angle and anterior-posterior center-of-rotation location (P < 0.001) than medial–lateral and anterior-posterior location (P = 0.0122). Individual variability accounted for a measurable proportion in variance explained of center-of-rotation location. Unique to gait, the lateral translation of center-of-rotation location resulted in the anterior translation of center-of-rotation at <10° knee flexion. Furthermore, no association between vertical ground-reaction force and center-of-rotation was identified.