With the rise of immersive visualization techniques, many domains within the manufacturing industry are increasingly validating production processes in virtual reality (VR). The validity of the results gathered in such simulations, however, is widely unknown-in particular with regard to human locomotion behavior. To bridge this gap, this paper presents an experiment, analyzing the behavioral disparity between human locomotion being performed without any equipment and in immersive virtual reality while wearing a head-mounted display (HMD). The presented study (n = 30) is split up in three sections and covers linear walking, non-linear walking and obstacle avoidance. Special care has been given to design the experiment so that findings are generally valid and can be applied to a wide range of domains beyond the manufacturing industry. The findings provide novel insights into the effect of immersive virtual reality on specific gait parameters. In total, a comprehensive sample of 18.09 km is analyzed. The results reveal that the HMD had a medium effect (up to 13%) on walking velocity, on non-linear walking towards an oriented target and on clearance distance. The overall-differences are modeled using multiple regression models, thus allowing the general usage within various domains. Summarizing, it can be concluded that VR can be used to analyze and plan human locomotion, however, specific details may have to be adjusted in order to transfer findings to the real world.