To date, limited work has been conducted investigating the biaxial elastic properties of the transverse carpal ligament (TCL), an important structure of the wrist with direct implications to carpal tunnel syndrome (CTS). Biaxial testing was conducted on human cadaveric TCL’s to assess differences in deep layer biaxial elastic moduli compared by region, direction and sex. Biaxial elastic moduli were found to be greater in the medial-lateral direction compared to the proximal-distal direction. Biaxial elastic moduli were also found to be greater in ulnar regions of the TCL compared to radial regions. The results of this work could be used in the development and/or improvement of computational and constitutive models of the TCL and help improve manipulative therapies for CTS.