Objective To separate less abundant proteins less than(10 000) Da with an effective Tricine-SDS-PAGE system.Methods The human serum albumin was removed by Hitrap Blue column,and serum was dealed with U9.Then gel was stained by the improved method(0.1% carbinol,0.5% TCA,0.1%Coomassie briliant blue G250).Results The discovery and detection of less abundant proteins less that(10 000) Da were improved obviously by removal of human serum albumin,serum treated with U9 and improvement of staining.Conclusion The improved Tricine-SDS-PAGE system is not only effective to resolve these less abundant proteins less than(10 000 Da,)but also favourable to recover these proteins from gel.