The aim of the study was functional evaluation of soft tissue mobilization in patients with temporomandibular disorder-myofascial pain with referral. The study group consisted of 50 individuals-37 females and 13 males. The average age was 23.36 ± 2.14 years. All subjects were diagnosed with myofascial pain with referral (diagnostic criteria for temporomandibular disorders). Soft tissue mobilization was applied three times. Electromyography of selected masticatory muscles was performed six times-before and after the treatment. After each mobilization, a decreasing tendency of muscular activity was observed in the entire study group. The Friedman test indicated that mobilization altered the activity of the right temporal muscle (p = 0.00010), both masseters (p = 0.0000), right sternocleidomastoid (p = 0.00251), left sternocleidomastoid (p = 0.00033), and right and left digastric muscles (p = 0.00045 and p = 0.00000, respectively). With respect to symmetry a statistically significant difference was noted in the case of the sternocleidomastoid muscles (p = 0.00729). In conclusion, soft tissue mobilization seems to be effective in the relaxation of masticatory muscles in patients with temporomandibular disorders. Our findings proved that soft tissue mobilization does not improve the symmetry and synergy of the masticatory muscles limited by dental occlusion.