Purpose Physical inactivity in conjunction with biological aging may lead to a decline in flexibility and neuromuscular function, contributing to physical frailty and an increased risk of injuries. Although, whole-body vibration has gained popularity as an alternative mode of exercise for improving physical fitness, only few studies examined the effects of WBV training on fitness parameters in middleaged individuals and the outcomes are not clear. This study investigated the effects of a 2-month whole-body vibration training (WBVT) program using a side-to-side vibration mode on flexibility, on power of lower limbs and on isokinetic peak torque of middle-aged women. Methods Twenty-four middle-aged untrained females (43.96 ± 3.28 years) were assigned to a vibration (WBVG) or a control group (CG). The WBVT program consisted of twenty-four sessions on a side-to-side vibration platform (frequency 20-25 Hz, amplitude 6 mm, duration 6-8 min). Flexibility, vertical jumping performance (VJ), and isokinetic peak torque of knee extensors (KE) and flexors (KF) were measured prior to and 2 days following the completion of WBVT. Results Post-training values of flexibility and VJ were significantly higher vs. pre-training values in WBVG (p \ 0.001-p \ 0.01); they remained unchanged in CG. Post-training flexibility and VJ values were significantly greater in WBVG vs. CG (p \ 0.001-p \ 0.05). The WBVT program had no effect on isokinetic peak torque of KE and KF. Conclusions An 8-week WBVT program improved flexibility and VJ in middle-aged untrained females and should be used by health and fitness professionals for counteracting the detrimental effects of sedentary lifestyle on flexibility and power of lower limbs.