The first objective of this study was to experimentally determine surface bone strain magnitudes and directions at the donor site for bone grafts, the site predisposed to stress fracture, the medial and cranial aspects of the transverse cross section corresponding to the stress fracture site, and the middle of the diaphysis of the humerus of a simplified in vitro laboratory preparation. The second objective was to determine whether computing strains solely in the direction of the longitudinal axis of the humerus in the mathematical model was inherently limited by comparing the strains measured along the longitudinal axis of the bone to the principal strain magnitudes and directions. The final objective was to determine whether the mathematical model formulated in Part I [Pollock, 2008, ASME J. Biomech. Eng., 130, p. 041006] is valid for determining the bone surface strains at the various locations on the humerus where experimentally measured longitudinal strains are comparable to principal strains. Triple rosette strain gauges were applied at four locations circumferentially on each of two cross sections of interest using a simplified in vitro laboratory preparation. The muscles included the biceps brachii muscle in addition to loaded shoulder muscles that were predicted active by the mathematical model. Strains from the middle grid of each rosette, aligned along the longitudinal axis of the humerus, were compared with calculated principal strain magnitudes and directions. The results indicated that calculating strains solely in the direction of the longitudinal axis is appropriate at six of eight locations. At the cranial and medial aspects of the middle of the diaphysis, the average minimum principal strain was not comparable to the average experimental longitudinal strain. Further analysis at the remaining six locations indicated that the mathematical model formulated in Part I predicts strains within standard deviations of experimental strains at four of these locations and predicts negligible strains at the remaining two locations, which is consistent with experimental strains. Experimentally determined longitudinal strains at the middle of the diaphysis of the humerus indicate that tensile strains occur at the cranial aspect and compressive strains occur at the caudal aspect while the horse is standing, which is useful for fracture fixation.