Matsumoto Y
Histochemistry. 1985;83(4):325-30. doi: 10.1007/BF00684378.
The use of the avidin-biotin technique in immunoperoxidase staining provides a simple and highly sensitive method for detecting the localization of antigens defined by monoclonal antibodies. However, endogenous biotin, which is widely distributed in tissues, often causes non-specific staining by binding to avidin [endogenous avidin-binding activity (EABA)]. Endogenous peroxidase activity (EPA) also makes the estimation of specific staining difficult. In the present study, several methods for the inhibition of EABA and/or EPA were examined using the avidin-biotin technique and monoclonal antibodies against murine Mac-1 and Ia antigen. Of these, the overnight incubation of sections in 40% methanol in phosphate-buffered saline containing 0.3% hydrogen peroxide gave the best result, as it inhibited EABA and EPA simultaneously without denaturating of the antigenic determinants recognized by the monoclonal antibodies.
抗生物素蛋白-生物素技术在免疫过氧化物酶染色中的应用为检测单克隆抗体所定义抗原的定位提供了一种简单且高度灵敏的方法。然而，广泛分布于组织中的内源性生物素常常通过与抗生物素蛋白结合（内源性抗生物素蛋白结合活性，EABA）而导致非特异性染色。内源性过氧化物酶活性（EPA）也使得特异性染色的评估变得困难。在本研究中，使用抗生物素蛋白-生物素技术以及针对小鼠Mac-1和Ia抗原的单克隆抗体，检测了几种抑制EABA和/或EPA的方法。其中，将切片在含0.3%过氧化氢的磷酸盐缓冲盐水中40%甲醇中过夜孵育效果最佳，因为它能同时抑制EABA和EPA，且不会使单克隆抗体识别的抗原决定簇变性。
