In order to study the proteome of Kandelia candel leaves, a two-dimensional electrophoresis (2-DE) system was established and optimized. Compared with the low protein content (1.7 μg μL-1) extracted by phenol (Phe), the improved trichloroacetic acid/acetone precipitation (TCA) method could obviously increase the extracting efficiency up to 12.4 μg μL-1. The protein spots of Kandelia candel leaves mainly distributed in the range of pH 4-7. The optimal loading quantity of samples were 400 μg gel-1 for a 17 cm IPG strip (pH 3-10 NL), which can detect 564±34 protein spots after silver staining. Therefore, the high resolution and repeatable 2-DE map could be obtained by using TCA extract method and the optimum system, which could widely use in proteome study of Kandelia candel.