Abstract: Objective To improve the in-gel digestion of 2-D gel electrophoresis after silver staining. Methods Modified method（adding non-ionic detergent n-dodecyl-β-D-maltoside in-gel digestion） and original method were compared by nanoUPLC-electrospray ionization tandem mass spectrometry（HD-MS）. Results Ten protein spots on 2-D gel were identified using the two methods.However,the amino acid sequence coverage in 60% proteins identified using the modified method was significantly higher than that identified using the original method. Conclusion The modified method is simple,and should be widely applied in study of gel-based proteomics.