Detection of protein blots using the avidin-biotin system.
Detection of protein blots using the avidin-biotin system.
复制标题
使用抗生物素蛋白-生物素系统检测蛋白质印迹。
DOI：
10.1385/0-89603-126-8:419
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发表时间：
1988
影响因子：
--
通讯作者：
K. Patel
中科院分区：
文献类型：
--
作者：
M. Dunn;K. Patel
聚丙烯酰胺凝胶电泳对复杂蛋白质混合物的高分辨能力只有在存在合适的程序来表征分离组分的情况下才能充分利用。解决这个问题的一种方法是研究分离的蛋白质与特定抗体或其他配体（如凝集素）的相互作用。这可以通过电泳后将抗血清直接应用于聚丙烯酰胺凝胶来实现，但这种“免疫固定”技术效率低且耗时，因为抗体扩散到凝胶基质中的速度很慢。这导致了将分离的蛋白质模式从凝胶转移到薄底物的方法的发展。这种技术被称为“Western”印迹。目前最流行的蛋白质底物是硝化纤维素。带正电的尼龙膜比硝化纤维素具有更大的蛋白质结合能力，可以成功地与抗体一起使用，但由于缺乏易于使用的通用染色程序，它们尚未被证明是流行的。
The high resolution capacity of polyacrylamide gel eletrophoresis for complex protein mixtures can only be fully exploited if suitable procedures exist for the characterization of the separated components. One approach to this problem is to investigate the interactions of the separated proteins with specific antibodies or other ligands such as lectins. This can be achieved by applying antisera directly to polyacrylamide gels after electrophoresis, but this technique of "immunofixation" is inefficient and time-consuming because of the slow rate of diffusion of antibodies into the gel matrix. This has resulted in the development of methods for transferring the pattern of separated proteins out of the gels onto thin substrates. This technique is known as "Western" blotting. The most popular substrate for proteins is currently nitrocellulose. Positively charged nylon membranes have a greater protein-binding capacity than nitrocellulose and can be used successfully with antibodies, but they have not proved popular because of the lack of an easy-to-use general staining procedure.