Objective: Altered body structures that occur with the loss of a lower limb can impact mobility and quality of life. Specifically, biomechanical changes that result from wearing a prosthesis have been associated with an increased risk of falls or joint degeneration, as well as increased energy demands. While previous studies describing these outcomes are typically limited by number of outcome measures and/or small, diverse patient groups, recent military conflicts present a unique opportunity to collect outcomes from a relatively homogenous, active patient population with limb loss. Thus, the objective of this study is to provide reference outcome measures on the basis of a large, relatively homogenous cohort of military personnel with transfemoral limb loss. Methods: A retrospective review of biomechanical, physiological, functional, and subjective measures was completed for 67 male servicemembers who sustained an injury resulting in traumatic, transfemoral limb loss during recent conflicts. These individuals represent a defined cohort, capable of exhibiting improved clinical outcomes resulting from demographic characteristics and extensive rehabilitation. Biomechanical and physiological outcome measures for 76 uninjured male servicemembers are also provided to serve as normative reference for full return to function. Select biomechanical and physiological outcomes related to stability, overuse, and efficiency are discussed in the text, on the basis of relevance to clinical gait assessment, in addition to functional and subjective measures. Results: In general, individuals with transfemoral limb loss exhibit decreased stability relative to uninjured individuals, noted by larger peak trunk velocity and step width variability; increased risk of low back and knee joint pain and/or degeneration, noted by larger trunk lateral flexion and bending moments, as well as larger vertical ground reaction force (vGRF) loading rates and impulses, respectively; and decreased efficiency during gait, noted by larger oxygen costs and leading limb mechanical work. Conclusion: Although the comprehensive set of measures presented here indicates overall reductions in biomechanical and functional performance with transfemoral limb loss compared to uninjured individuals, these reductions were relatively smaller than existing evidence among populations that are more diverse in age and activity level. Therefore, this data set may be used as benchmarks for young, active individuals with transfemoral limb loss, to assist with setting clinical goals, and to aid in the evaluation of new treatment techniques or interventions. These measures will also be particularly important for subsequent evaluations and longitudinal follow-ups to determine the longer-term impact of transfemoral limb loss on this cohort.