Trojanowski J Q, Obrocka M A, Lee V M
J Histochem Cytochem. 1983 Oct;31(10):1217-23. doi: 10.1177/31.10.6350434.
Eight different previously described chromogen protocols were evaluated with respect to their sensitivity for the visualization of horseradish peroxidase (HRP) in a peroxidase-antiperoxidase (PAP) complex used with the unlabeled antibody method for immunohistochemistry. The protocols were evaluated in a test system that involved the demonstration of immunoreactive neurofilaments (NF) or glial filaments (GF) in paraffin-embedded sections of rat cerebellum using anti-NF or anti-GF monoclonal antibodies (MA). The chromogens included: amino-ethylcarbazole (AEC), diaminobenzidine (DAB), O-tolidine, paraphenylenediamine-pyrocatechol (PPD-PC), and tetramethylbenzidine (TMB). The incubation medium using DAB as the chromogen was employed at neutral pH, at pH 5.1, or at neutral pH with the addition of either cobalt chloride or imidazole to intensify the reaction product. The relative sensitivity of the chromogen protocols was quantitated by comparing the dilution of the anti-NF or anti-GF MA at which NF or GF immunoreactivity was extinguished using each protocol. The results obtained with both the anti-NF and anti-GF MA indicated that DAB with imidazole was the most sensitive chromogen protocol.
针对免疫组织化学中未标记抗体法所使用的过氧化物酶 - 抗过氧化物酶（PAP）复合物中辣根过氧化物酶（HRP）可视化的灵敏度，对之前描述的8种不同的显色剂方案进行了评估。这些方案在一个测试系统中进行评估，该系统涉及使用抗神经丝蛋白（NF）或抗胶质纤维（GF）单克隆抗体（MA），在大鼠小脑石蜡包埋切片中显示免疫反应性神经丝（NF）或胶质纤维（GF）。显色剂包括：氨基乙基咔唑（AEC）、二氨基联苯胺（DAB）、邻联甲苯胺、对苯二胺 - 邻苯二酚（PPD - PC）和四甲基联苯胺（TMB）。以DAB作为显色剂的孵育介质在中性pH、pH 5.1或中性pH并添加氯化钴或咪唑以增强反应产物的条件下使用。通过比较使用每种方案使NF或GF免疫反应性消失时抗NF或抗GF MA的稀释度，对显色剂方案的相对灵敏度进行定量。使用抗NF和抗GF MA获得的结果表明，添加咪唑的DAB是最灵敏的显色剂方案。
