[Aim] Protein sample preparation is the prerequisite to obtain an optimal two-dimensional electrophoresis (2-DE) gel map. Establishing a suitable 2-DE system can provide a strong support for the subsequent research.[Method] This study used Frankliniella occidentalis adults as experimental materials, and compared three protein extraction methods:Tris-phenol, TCA/acetone precipitation, and direct decomposition extraction. The most suitable method of 2-DE system was defined through the analysis of 2-DE protein images.[Result] Experimental results revealed that the three methods showed significant differences in protein extraction rate, among which the direct decomposition method was the highest while Tris-phenol method was the lowest. The three methods got similar SDS-PAGE results. TCA/acetone method had the best quality of 2-DE gel map, with the highest number of protein spots.[Conclusion] The results suggest that TCA/acetone method could remove interfering substance effectively and was optimal for protein extraction of F. Occidentalis. It laid foundation of further study on the proteomics of F. occidentalis.