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尿液标本的双向电泳技术优化

DOI: 10.13604/j.cnki.46-1064/r.2015.09.03, PP. 1040-1043

Keywords: 尿液,双向电泳,超滤沉淀法,蛋白组学

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Abstract:

目的通过摸索尿液的蛋白制备的实验步骤,总结一套适用于尿液的蛋白制备方案及其双向凝胶电泳技术,提高尿液标本2Dgel(Two-dimensionalpolyacrylamidege,双向电泳胶)的质量。方法对双向电泳实验中的关键环节如蛋白的沉淀,高峰度蛋白的去除等进行实验优化,热考马斯亮蓝染色后进行凝胶图像比较。结果采用超滤法沉淀蛋白,并结合血清亲和法试剂盒(AlbuminandIgGremovekit)去除高峰度蛋白,可以得到分辨率高、约700个蛋白点数的2D胶,质谱鉴定出Tubulinalphachain-like3及Tropomyosinbetachain两个蛋白。结论对超滤沉淀后的蛋白,应用血清亲和法试剂盒(AlbuminandIgGremovekit),去除尿液高峰度蛋白,可大大提高尿液标本2D胶的质量,质谱鉴定结果也提示所分离的蛋白与肾脏疾病相关。以上为肾脏疾病蛋白质组学研究奠定基础。

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