Several studies have shown that eggshell membrane (ESM) is a suitable biomaterial with potential applications in biomedicine such as wound repair and cell culture. In order to control and improve the use of ESM for biomedical applications their physical and structural properties must be known. In this paper, we have studied the effect of temperature on the mechanical properties of the ESM. Atomic force microscopy was used to assess the morphology of the ESM. The mechanical properties of the membranes were studied by means of uniaxial tensile tests carried out at four different temperatures. Differential scanning calorimetry and thermo-gravimetrical analysis were used to assess the thermal transitions of the ESM and the influence of the water content on its thermal behavior. The Young's modulus showed a linear inverse dependence with regard to the temperature of the sample. A peak associated to the thermal denaturation of collagen was observed in the DSC tests of the membrane. These peaks showed a dependence on the water content of the specimens. © 2013 Akadémiai Kiadó, Budapest, Hungary.