血清样本双向电泳实验方法学研究

目的提高血清双向电泳实验的分辨率,建立理想的血清双向电泳实验方法。方法对血清双向电泳(two-diensional gel electrophoresis,2-DE)中血清标本的处理方法以及硝酸银染色方法等条件和技术方法进行了调整和优化。结果经过优化后,建立了重复性较好的血清2-DE实验方法 ,血清样本中蛋白质斑点的分辨率得到了明显的提高,蛋白质斑点明显增多,且此方法能够兼容后续生物质谱分析。结论通过优化和验证,建立了较实用的血清2-DE实验方案,对血清样本的蛋白质组学分析具有很好的技术参考价值,为进一步分析、寻找和鉴定药物干预或与疾病相关的血清蛋白质奠定了实验基础。

Method studies of two-dimensional polyacrylamide gel electrophoresis of serum

Aim To establish an ideal two-dimensional polyacrylamide gel electrophoresis( 2-DE) method for serum proteomic research. Methods The 2-DE experiment of serum was opti-mized by adjusting the conditions of sample pretreatment and silver nitrate staining. Results An ideal 2-DE experiment method with higher repetition was established. The number of protein spots and the resolution were both increased,and this method was compatible with MS analysis. Conclusion After the adjustments and optimizations,an ideal method of 2-DE technique for serum is established,which has a high reference value for the relevant researches,and lays an experimental foundation for the search,analysis as well as identification of the disease related proteins in serum.