胃粘膜组织蛋白质组双向凝胶电泳技术的初步建立与优化

PRELIMINARY ESTABLISHMENT AND OPTIMIZATION OF TWO-DIMENSIONAL GEL ELECTROPHORESIS FOR PROTEOMICS OF GASTRIC MUCOSA

为建立并优化人类胃粘膜组织蛋白质组分析所需的双向凝胶电泳技术 ,提高其分辨率及重复性。采用刮取手术胃粘膜组织 ,对以固相pH梯度为第一向的双向凝胶电泳的关键因素与环节 ,如样品处理、上样量、电泳参数、凝胶浓度和SDS凝胶电泳染色方法等进行一系列的优化。结果以固相pH梯度———IPG胶条 (pH =3～10 )进行第一向等电聚焦 ,以SDS均一胶 (13% )的垂直电泳为第二向 。

Two dimensional gel electrophoresis(2 DE) is a key technique for proteomics. To analyze the proteome of gastric mucosa and its changes in different diseases. 2 DE technique was established. A series of important factors, such as sample preparation, electrophoresis parameters, choice of IPG gels, concentration of SDS gels, and protocol for staining were optimized to improve the resolution and reproducibility. By using the proper methods describeds above, choosing appropriate sample amount, using pre cast IPG dry strip and casting 13% equal gel, satisfactory 2 DE maps of gastric mucosa were obtained.