In the beamhouse, liming might directly affect the structure and performance of collagen, as well as the quality of resultant leather. However, the influences of composition and content of liming agents on liming mechanism are quite complicated. In this study, calcium hydroxide and urea were utilized in liming. The solution pH, hide swelling ratio and non-collagenous protein removal were quantitatively analyzed. The morphologies of both limed and fresh hides were studied by optical microscopy. The reaction mechanism of fiber opening up of cattle hides was analyzed and speculated by the combination of thermogravimetric analysis and Fourier transformation infrared spectroscopy. It was found that the fiber bundles of hides limed by urea/calcium hydroxide have a better opening up effect than that by pure calcium hydroxide. The mechanism of liming in an urea/calcium hydroxide solution system was proposed.