Abstract: Sample preparation is the key to the success of proteomics studies.To obtain a high-quality electrophorectogram,a two-dimensional electrophoresist was established which was suitable for the proteomic analysis of ramie's various parts(stems,leave) by optimizing the traditional TCA/acetone protein extraction method.An improved 2-DE technology system was obtained:Tris and Triton X-100 were added together instead of CHAPS to the lysis buffer,protein purification (4 times volume of acetone) after protein cleavage,IPG strips (pH=4—7,24cm),loading amount was 800μg/strip(Coomassie Brilliant Blue stain).The method effectively eliminated nonprotein impurities and obtained a clear two-dimensional gel electrophoresis array. The protein of hemp was found mainly distributed at the range of pH=5—6,the protein spots of stem and leaves were 1027±12,905±8 respectively.This technology was also applied to the stems and leaves of hemp and provided reliable technical support to the proteomic study of hemp.