Background: Iliotibial band syndrome (ITBS) represents one of the most common running related injuries. The pathophysiology is postulated to be caused by excessive ITB tension, impingement and irritation of soft tissues at the lateral femoral epicondyle. However, direct evidence has yet to be found and the multifactorial etiology is under discussion. The purpose was to evaluate stiffness of ITB, gluteus maximus (GM) and tensor fasciae latae (TFL) muscles using shear wave elastography (SWE). Methods: In 14 patients with clinically verified ITBS and 14 healthy controls, three SWE measurements each of ITB, GM and TFL in both legs was performed to determine measurement reliability and between-group and -leg differences. Results: The mean value of ITB was 12.8 m/s with ICC of 0.76, whereas the values measured in the GM were 3.02 m/s with an ICC of 0.87. No statistically significant difference in controls compared to patients were found (p = 0.62). The mean value of TFL was 5.42 m/s in healthy participants, compared to 3.89 m/s patients with an ICC of 0.98 (p = 0.002). Conclusion: Although SWE showed no difference in ITB stiffness, significant differences for TFL muscle stiffness in runner’s knee was found, suggesting that the hip abductor muscles might play a bigger role in the pathophysiology of ITBS. We aimed to implement baseline values for stiffness assessments and prove reliability for further prospective studies of SWE in runner’s knee.