De Jong A S, Van Kessel-van Vark M, Raap A K
Histochem J. 1985 Oct;17(10):1119-30. doi: 10.1007/BF01002537.
Various chromogen protocols for visualizing peroxidase and alkaline phosphatase activity in immunoenzyme histochemistry were compared with respect to their sensitivity. They were tested on tissue sections of human skeletal muscle and in an antigen spot test using antibodies against slow skeletal muscle myosin. The chromogens included 3-amino-9-ethylcarbazole (AEC), 3, 3'-diaminobenzidine (DAB), p-phenylenediamine-pyrocatechol (PPD-PC) and 4-chloro-1-naphthol (CN) in peroxidase histochemistry, and 5-bromo-4-chloro-3-indolyl phosphate-nitro blue tetrazolium salt (BCIP-NBT), BCIP-tetra nitro blue tetrazolium salt (TNBT) and various combinations of substituted naphthol phosphate-diazonium salt in alkaline phosphatase histochemistry. DAB, CN, and PPD-PC were also employed with imidazole and DAB in addition to Co2+ and Ni2+ ions. The results indicate that DAB-imidazole and DAB-Co2+ and Ni2+ ions are the most sensitive chromogen protocols for visualizing peroxidase activity. Although no large differences were found between the various chromogen protocols for visualizing alkaline phosphatase activity, the protocol BCIP-TNBT is especially recommended. Furthermore, the various chromogen protocols were evaluated as to stability of chromogen solutions and final precipitates, background staining, localization properties, and enhancement of enzyme activity.
在免疫酶组织化学中，就其敏感性对用于可视化过氧化物酶和碱性磷酸酶活性的各种显色剂方案进行了比较。在人类骨骼肌组织切片上以及使用抗慢肌骨骼肌肌球蛋白抗体的抗原斑点试验中对它们进行了测试。过氧化物酶组织化学中的显色剂包括3-氨基-9-乙基咔唑（AEC）、3,3'-二氨基联苯胺（DAB）、对苯二胺-邻苯二酚（PPD-PC）和4-氯-1-萘酚（CN），碱性磷酸酶组织化学中的显色剂包括5-溴-4-氯-3-吲哚基磷酸-硝基蓝四唑盐（BCIP-NBT）、BCIP-四硝基蓝四唑盐（TNBT）以及取代萘酚磷酸-重氮盐的各种组合。DAB、CN和PPD-PC还与咪唑以及除了Co2+和Ni2+离子之外的DAB一起使用。结果表明，DAB-咪唑以及DAB-Co2+和Ni2+离子是用于可视化过氧化物酶活性最敏感的显色剂方案。尽管在用于可视化碱性磷酸酶活性的各种显色剂方案之间未发现大的差异，但特别推荐BCIP-TNBT方案。此外，就显色剂溶液和最终沉淀物的稳定性、背景染色、定位特性以及酶活性增强对各种显色剂方案进行了评估。
