Abstract: Using the hepatopancreas of Jade perch as the experimental samples, the tissue lysate formula and ratio of lysis buffer volume to tissue weight, as well as the isoelectric focusing electrophoresis procedures were screened. The results were as follows: lysate formula I（8 M Urea, 2 M Thiourea, 0.5%（W:V）of CHAPS, 40 mM Tris-Base, 1% DTT, 2%（W:V）Pharmalyte pH 3-10, 1 mM PMSF）was the better. When the ratio Lysate volume to the sample weight was 1:3, the lytic effect was the better. The electrophoretic program І（S1:0-500 V, 500 V•h, S2:500 V, 500 V•h, S3:500-3 500 V, 10 000 V•h, S4:3 500 V, 50 000 V•h. S5:3 500-5 000 V, 8 000V•h）had the better isoelectric focused effect. Based on these results, a protocol of two-dimensional gel electrophoresis（2-DE）was set up. The optimized 2-DE techniques provided a basis for studying the proteome of Jade perch hepatopancreas.

Key words: Jade perch（Scortum Barcoo）, hepatopancreas, proteome, 2-DE