Abstract: Peach （Prunus persica L.‘Okubao’）fruit was adopted in this experiment. Comparing different effects of two-dimensional electrophoresis （2-DE）gels that were involved with quantitative loading of sample, sample buffer, equilibration buffer and the type of staining to explore influence factors of 2-DE patterns on peach fruit. Although the two-dimensional gel stained by coomassie brilliant blue had gained less protein spots than the gel stained by silver, it had a clear background intensity and was applicable to the proteomic study. Furthermore, approximately 100 μg of protein was brought to a final volume of 375 μL sample buffer which contained thiourea and sulfphobetaine 3-10 and loaded on 17 cm immobilized pH gradients strip, then the strip was incubated in equilibration buffer with tributyl phosphine after isoelectric focusing. The two-dimensional gel stained by silver detected 1 209 protein spots and had slight streaks. The results demonstrated that the protocol was suited to proteomic analysis of peach fruit and compatibled with protein identification by mass spectrometry of downstream processing.

Key words: Prunus persica, Fruit, Protein, Two-dimensional electrophoresis