The mechanical properties of the transverse carpal ligament (TCL) have yet to be comprehensively measured. Failure strain, failure load, ultimate strength, stiffness and elastic modulus were reported from uniaxial tensile tests. On human cadaveric TCLs, failure strain was significantly higher in female specimens compared to males. Moreover, regional strain analysis showed the lateral end of the tissue to strain significantly more than the medial. Biaxial testing at three different tissue depths was also carried out. Mechanical properties were found to vary significantly by depth with the deeper layers displaying higher mechanical properties. A significant difference with respect to loading direction was also observed with the medial/lateral direction displaying higher mechanical properties than proximal/distal. These results provide mechanical properties which can be incorporated into finite element models or constitutive models of the TCL. Furthermore, this work provides a basis for recommendations for noninvasive treatment options for carpal tunnel syndrome.