Porter D D, Porter H G
J Immunol Methods. 1984 Aug 3;72(1):1-9. doi: 10.1016/0022-1759(84)90428-9.
Separation of mixtures of proteins by polyacrylamide gel electrophoresis followed by transfer of the proteins to support media such as nitrocellulose and detection by immunologic procedures provides a powerful analytic tool for assaying the antibody specificity of antisera or for following the purification of antigens. This technique requires fewer assumptions about antigenic solubility or antibody reactivity than immunoprecipitation methods. We present a glucose oxidase immunoenzyme staining procedure for protein blots and illustrate its use for detecting antibody to several viruses. The glucose oxidase immunoenzyme stain has a lower background than some peroxidase stains. We have detected as little as 1 microgram/ml of antiviral antibody using this stain.
通过聚丙烯酰胺凝胶电泳分离蛋白质混合物,随后将蛋白质转移至诸如硝酸纤维素等支持介质上,并通过免疫程序进行检测,这为测定抗血清的抗体特异性或追踪抗原的纯化提供了一种强大的分析工具。与免疫沉淀方法相比,该技术对抗抗原溶解度或抗体反应性的假设更少。我们提出了一种用于蛋白质印迹的葡萄糖氧化酶免疫酶染色程序,并举例说明了其用于检测几种病毒抗体的用途。葡萄糖氧化酶免疫酶染色的背景比一些过氧化物酶染色更低。使用这种染色方法,我们已检测到低至1微克/毫升的抗病毒抗体。