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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 gel electrophoresis for proteome analysis of human testicular tissue and improve its resolution and repeatability. The method was separated by two-dimensional gel electrophoresis (SDS-PAGE). The gel was stained by silver staining and analyzed by 2-DE with Melanie 4 software. The sample processing, sample loading, strip selection and other steps were a series of optimization. The results obtained successfully obtained human testis tissue with high resolution, good reproducibility of the two-dimensional electrophoresis. Conclusion The proteome map of human testis was established by two-dimensional gel electrophoresis, which laid the foundation for the further study on proteomics of related human diseases.