The measurement of gait parameters normally requires motion tracking systems combined with force plates, which limits the measurement to laboratory settings. In some recent studies, the possibility of using the portable, low cost, and marker-less Microsoft Kinect™ sensor to measure gait parameters on over-ground walking has been examined. The current study further examined the accuracy level of the Kinect sensor for assessment of various gait parameters during treadmill walking under different walking speeds. Twenty healthy participants walked on the treadmill and their full body kinematics data were measured by a Kinect sensor and a motion tracking system, concurrently. Spatiotemporal gait parameters and knee and hip joint angles were extracted from the two devices and were compared. The results showed that the accuracy levels when using the Kinect sensor varied across the gait parameters. Average heel strike frame errors were 0.18 and 0.30 frames for the right and left foot, respectively, while average toe off frame errors were −2.25 and −2.61 frames, respectively, across all participants and all walking speeds. The temporal gait parameters based purely on heel strike have less error than the temporal gait parameters based on toe off. The Kinect sensor can follow the trend of the joint trajectories for the knee and hip joints, though there was substantial error in magnitudes. The walking speed was also found to significantly affect the identified timing of toe off. The results of the study suggest that the Kinect sensor may be used as an alternative device to measure some gait parameters for treadmill walking, depending on the desired accuracy level.