Abstract: This paper reported a improvement in 2-D gel electrophoresis of the proteome in female cones of the female-sterile Pinus tabulaeformis Carr. An IPGphorTM unit with immobile pH gradient strips was used in and the EttanTM DALT Ⅱ system under constant power and temperature in SDS-PAGE. The 2-D gel was stained by both methods of coomassie blue and silver.Extraction of proteins, pH of the strips, rehydration of the strips and other parameters were optimized and compared. Higher repeatability and better separating protein pattern could be gained by this technique.