Abstract: The methods of extracting proteins from ovules of Pinus tabulaeformis Carr. were compared and optimized in this paper. The buffers of 0.05 mol/L Tris-HCl containing two different SDS concentrations, four different pH values and a TCA/acetone solution were used.Resultsindicated that the effects of the pH value of the Tris-HCl buffer on SDS-PAGE were relevant to the SDS concentration and the best result occurred at pH 8.8 by using a buffer containing 0.2% SDS. While using the buffer containing 2% SDS, there were no significant differences among different pH values, and protein bands of 14.4, 16.1 and 17.1 kD were obviously strengthened. For IEF/SDS-PAGE, the TCA/acetone precipitation method was the best since the 2-D map was clearer and there were more protein spots than the others. However, among three Tris-HCl buffers, the best 2-D pattern was obtained by using a Tris-HCl buffer (pH 8.8) containing 0.2% SDS.