Optimization method for the extraction and separation of proteins from ovules of Pinus tabulaeformis Carr

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.