To find out a suitable method for refining protein of Jinhua ham muscle tissue,lysate method,TCA∕acetone precipitation method,2-DE clean-up kit,TCA∕acetone and clean-up kit,gel column chromatography method were used.In the 2-DE system,17 cm pI 5～8 IPG gel was chosen as the first dimension of isoelectro-focusing electrophoresis;the second dimension was SDS-polyacrylamide gel electrophoresis,300ug protein was segregated and the 2-DE maps were obtained.The electropherogram were analyzed by using image analysis software.The Results showed the horizontal stripes,vertical stripes,and background of the 2-DE maps of protein refined by gel column chromatography method decreased obviously.The protein spots obtained by gel column chromatography method increased significantly.At the same time the samples obtained by gel column chromatography method were more stable in isoelectric focusing.This may provided a foundation for the further study of the Jinhua ham characteristic protein and its changes.