The purpose of this study was to identify side-to-side and sex-based differences in hip kinematics during a unilateral step-up from deep flexion. Twelve (eight men, four women) asymptomatic young adults performed a step ascent motion while synchronized biplane radiographs of the hip were collected at 50 images per second. Femur and pelvis position were determined using a validated volumetric model-based tracking technique that matched digitally reconstructed radiographs created from subject-specific computed tomography (CT) bone models to each pair of synchronized radiographs. Hip kinematics and side-to-side differences were calculated and a linear mixed effects model evaluated sex-based differences. Women were on average 10.2 deg more abducted and 0.2 mm more medially translated than men across the step up motion (p < 0.001). Asymmetry between hips was up to 14.1 ± 12.1 deg in internal rotation and 1.3 ± 1.4 mm in translation. This dataset demonstrates the inherent asymmetry during movements involving unilateral hip extension from deep flexion and may be used provide context for observed kinematics differences following surgery or rehabilitation. Previously reported kinematic differences between total hip arthroplasty and contralateral hips may be well within the natural side-to-side differences that exist in asymptomatic native hips.