中枢神经蛋白质组分析中双向电泳技术的建立

建立和优化了中枢神经组织蛋白质组分析所需的双向电泳及相关技术.由于中枢神经组织结构的特殊性，样品处理非常困难.对样品液组成、样品处理、上样方式、上样量、IPG胶条和SDS-聚丙烯酰胺凝胶电泳染色方法和保存等相关技术进行了比较研究和条件优化后，以固相pH梯度等电聚焦为第一向和SDS均一胶（T=12.5%）的水平电泳为第二向，成功地得到了神经组织双向电泳图谱.

Improvement of Two-dimensional Gel Electrophoresis for Proteomics of Rat Central Nervous System

Two-dimensional gel electrophoresis (2-DE) is a key technique for proteomics. To analyze the proteome of PC12 cells and rat central nerve tissues, including brain and spinal cord, 2-DE technique is established. Due to much lipid and other non-protein interfering constituents, protein extraction is much more difficult for nerve tissues than for other tissues. Two different methods, i.e., precipitation with tricholoroacetic acid/acetone and ultra-centrifugation were employed to extract protein from rat brain and spinal cord for 2-DE. Other factors such as method and volume of loading sample, choice of IPG gels, concentration of SDS gels, preset of electric parameters, protocol for staining and drying the gels were also improved. Using the proper method described above, satisfactory 2-DE maps of PC12 cells, rat brain and spinal cord were obtained.