目的:建立和优化人表皮组织蛋白质组双向电泳技术,为后续的皮肤蛋白质组学研究奠定基础。方法:组织匀浆法制备表皮组织总蛋白,采用固相pH梯度胶条进行双向电泳,凝胶银染后用PDQuest7.4软件对电泳图谱进行分析。结果:人表皮组织蛋白质组在7cm pH5~8 IPG胶条的最佳上样量为200μg,平均蛋白斑点数为414±19,平均匹配斑点数为346±31,匹配率达83.6%。结论:通过优化双向电泳条件,获得了分辨率高、重复性好的人表皮组织蛋白质组双向电泳图谱。
Aim: To establish two-dimensional electrophoresis methods for human epidermis proteome. Methods: The total proteins of human epidermis were extracted by homogenization and separated by immobilized pH gradient (IPG) strips prior to SDS polyacrylamide gel electrophoresis. The gels were stained with silver nitrate, scanned and then analyzed using PDQuest 7.4 analysis software. Results: With 200 μg of protein loaded on 7 cm pH 5 ~ 8 IPG strips, 414 ± 19 protein spots were detected in the two-dimensional electrophoresis profiles. The matched spots between gels were 346 ± 31 and the match rate was 83.6%. Conclusion :The two-dimensional electrophoresis profiles of human epidermis proteome were obtained with high resolution and reproducibility.