摘要
应用不同蛋白质复性剂、封闭剂和标记物的对比方法以探讨Ｗｅｓｔｅｒｎ印迹技术的改进。结果表明：①经 （ＴｒｉｔｏｎＸ－１００）＝０．３％，５ｍｉｎ和ＳＢＢ２０ｍｉｎ的蛋白质复性处理后，可使第一免疫血清稀释度１：１００改为１：１０００使用；②应用 （Ｔｗｅｅｎ２０）＝０．０５％作为封闭液效果最好；③金标法及酶标法都可用于Ｗｅｓｔｅｒｎ印迹技术，酶标法主带清晰，而金标法灵敏度高。
The effects of different renaturation agents, blockings and labelled conjugates were studied to improve the technique of non-radioactive Western Blot.The results showed that ①after the treatment with 0. 3% Triton X-100 5 min or standard binding buffer (SBB) 20 min,the proteins on NCM could be detected with first specific antiserum from the titer 1:100 diluted to 1: 1000; ②the blocking effect of Tween 20 was more effective than that of other blockings; ③both HRP-labelled conjugate and colloid gold-labelled conjugate could be used as the labelled conjugate in Western Blot. The main band detected with HRP-labelled conjugate was clear on NCM, while the colloid gold-labelled conjugate was much more sensitive than the HRP-labeled one.
出处
《河南医科大学学报》
1996年第1期10-12,共3页
Journal of Henan Medical University
基金
国家自然科学基金
