Bindl J M, Warnke R A
Am J Clin Pathol. 1986 Apr;85(4):490-3. doi: 10.1093/ajcp/85.4.490.
We describe a method of biotin/avidin-peroxidase detection using second and third stage reagents in Coplin jars. This method allows a large quantity of sections to be stained simultaneously with a minimal amount of technical time involved. A wide range of mouse monoclonal antibodies of varying specificities and isotypes were used to stain both frozen and paraffin-embedded sections of various normal and neoplastic tissues. Three different biotinylated anti-mouse antibodies were tested, including F(ab')2 antibody fragments of one, followed by horseradish peroxidase conjugated avidin. All monoclonal antibodies employed gave good staining, using incubation times of 30-50 minutes. The staining was done during a mean period of 25 to 27 days with an average staining load of 500 sections per Coplin jar.
我们描述了一种在 Coplin 罐中使用第二和第三阶段试剂进行生物素/抗生物素蛋白-过氧化物酶检测的方法。该方法允许同时对大量切片进行染色,且所需技术时间最少。使用了多种具有不同特异性和亚型的小鼠单克隆抗体对各种正常和肿瘤组织的冰冻切片及石蜡包埋切片进行染色。测试了三种不同的生物素化抗小鼠抗体,其中一种是 F(ab')2 抗体片段,随后使用辣根过氧化物酶偶联抗生物素蛋白。所有使用的单克隆抗体在 30 - 50 分钟的孵育时间内都能产生良好的染色效果。染色在平均 25 至 27 天的时间内完成,每个 Coplin 罐平均染色量为 500 个切片。