摘要
目的建立并优化人睾丸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。方法利用固相pH梯度双向凝胶电泳技术进行分离,凝胶经银染显色后,用Melanie 4软件分析2-DE图谱。对样品处理、上样量,胶条的选择等步骤进行了一系列的优化。结果优化后成功地获得了人睾丸组织分辨率高、重复性好的双向电泳图谱。结论应用双向凝胶电泳技术建立了人睾丸组织蛋白质组图谱,为其及相关疾病的蛋白质组学的进一步研究奠定了基础。
Objective To establish and optimize two-dimensional electrophoresis（2-DE） for proteome analysis of the human testicular tissue and to promote resolution and reproducibility. Methods Immob＇dized pH gradient two-dimensional polyacrylamide gel electrophoresis was used to separate the total proteins of the samples, and protein spots separated were dyed by silver staining. Protein profile were analyzed by melanie 4 image analysis software.Several key steps related 2DE such as sample preparation, loading amount, choice of 1PG strips etc,were opotimized. Results A high resolution and reproducible 2-DE image of the human testicular tissue is successfully obtained after optimizing. Conclusion Establishment of ideal proteome profile based on 2-DE was the premise for comparative proteomics study of diseases.
出处
《中国男科学杂志》
CAS
CSCD
2009年第4期3-6,共4页
Chinese Journal of Andrology
基金
国家自然基金(30772167)
广州市科技计划项目(2008J1-C131)
关键词
电泳
凝胶
双向
蛋白质组
睾丸
electrophoresis, gel, two-dimensional
proteome
testis
