非离子去污剂提高双向电泳银染后胶内酶切的效率

Non-ionic detergent n-dodecyl-β-D-maltoside improves the in-gel digestion of 2-D gel electrophoresis after silver staining

目的改进二维凝胶电泳银染色后的胶内酶切方法,提高蛋白质的鉴定率。方法应用超高效纳升液相色谱-电喷雾串联质谱(HD-MS),对改进方法 (酶切中加入非离子去污剂正十二烷-β-D-麦芽糖苷)和原有方法进行比较。结果 2D胶上的10个蛋白质斑点中的蛋白质用两种方法均得到明确的鉴定结果,但60%的蛋白质用改进方法鉴定的氨基酸序列覆盖率较原有方法明显提高。结论改进后的方法简便,适用于凝胶为基础的蛋白质组学研究。

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.