A 2-DE (two-dimensional electrophoresis) protocol for total proteins of Acacia implexa Benth. was described. The yield of water-soluble proteins was increased if 10% PVP was added in the protein extraction buffer, centrifugal force was increased and centrifugal time was prolonged. The optimum loading quantity of sample proteins was 1 000 μg in isoelectric focusing. The optimum concentration of SDS--polyacrylamide gel electrophoresis was 12.5 %. The optimized protocol markedly minimized the interference from wood pigments and other non-protein substances．