Abstract:An effective protein extraction method is crucial to separate the proteins by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Tung tree (Vernicia fordii) seed, which is used for producing the tung oil, contains large amounts of compounds which can interfere with the extraction of proteins, and can even affect the resolution of 2D-PAGE images. In the present study, we developed a suitable protein extraction method from three tested methods, including trichloroacetic acid (TCA)-acetone (Method A), phenol (Method B), TCA-acetone combined with phenol (Method C). The results showed that the protein concentration of samples by Method C was high and it achieved 8.1 μg·μL-1. Method C has a strong capability to extract the high-molecular-weight and the low-molecular-weight proteins from tung tree seeds. In addition, the 2D-PAGE image resolution of Method C was also better than the other two methods. Thus, Method C was used to extract proteins, and it would lay the foundation for proteomic analysis on the tung tree seeds.