Whole body vibration (WBV) exercise has been introduced into human and recently also into equine training. Only a few studies about physical and physiological effects on horses are available. This study should clarify the actual physical vibration of a commercially available WBV plate itself and on the horse as well as the muscular activity in the limbs and back. Furthermore, the effects of WBV warm-up on clinical parameters and body surface temperatures were compared to standard warm-up exercises. Ten sound horses (vibration and muscle activity of six horses) were recorded while standing (control) and during 15 and 25 Hz (manufacturer information) vibration exercise. The vibration of plate, hoof, fetlock, withers, and sacrum was analyzed for frequency, peak-to-peak displacement, and peak acceleration. Activity of M. triceps, quadriceps, and longissimus dorsi was assessed using surface electromyography. Warm-up effects were compared between four different warm-up scenarios: standing (control), 10-minute vibration, 10-minute lunging (walk), and 12-minute lunging (walk and trot). Maximal body surface temperature of upper forelimb, thigh, and back was measured. Actual plate vibration frequency was 7 or 11 Hz with a maximum peak-to-peak displacement of 9 mm in longitudinal direction. WBV exercise induced no increase in electromyographic activity, clinical parameter, or body surface temperature. It was concluded that actual plate vibration was mainly longitudinal with a lower frequency than proposed and 10-minute exercise had no significant effect. Different vibration protocols and vibration acting in the vertical direction might enable more effective exercise in horses.