A NEW STABILIZING AGENT FOR THE TETRAMETHYL BENZIDINE (TMB) REACTION-PRODUCT IN THE HISTOCHEMICAL DETECTION OF HORSERADISH-PEROXIDASE (HRP)
A NEW STABILIZING AGENT FOR THE TETRAMETHYL BENZIDINE (TMB) REACTION-PRODUCT IN THE HISTOCHEMICAL DETECTION OF HORSERADISH-PEROXIDASE (HRP)
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DOI：
10.1016/0165-0270(85)90025-1
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发表时间：
1985-01-01
影响因子：
3
通讯作者：
LOPEZGARCIA, C
中科院分区：
文献类型：
--
作者：
OLUCHA, F;MARTINEZGARCIA, F;LOPEZGARCIA, C
本文报道了一种以七钼酸铵（AHM）为稳定剂，四甲基联苯胺（TMB）为显色剂的辣根过氧化物酶（HRP）组织化学检测方法。该方法避免了使用硝铁氰化钠（SNF）作为稳定剂的先前方法中出现的2个主要问题，即，在非特异性解剖部位出现针状晶体和强烈的组织收缩。比较研究了TMB-AHM和TMB-SNF两种方法在分析蜥蜴Podarcis hispanica大脑皮层传入联系中的作用。两种方法的灵敏度相似。TMB-AHM反应可以在生理pH（6-8）下进行，从而避免组织收缩。反应产物为强烈的蓝绿色，并且与TMB-SNF方法一样，显示出造粒。当孵育介质保持在pH 5时，完全避免了非特异性沉淀物的出现。
An alternative procedure for the histochemical detection of HRP [horseradish peroxidase] using ammonium heptamolybdate (AHM) as a stabilizing agent and tetramethyl benzidine (TMB) as a chromogen is reported. This procedure avoids the 2 main problems that occur in previous methods using sodium nitroferricyanide (SNF) as the stabilizer, namely, the appearance of needle-shaped crystals at non-specific anatomical sites and intensive tissue shrinkage. A comparative study of both, the TMB-AHM and TMB-SNF methods, was performed in the analysis of cerebral cortex afferent connections of the lizard Podarcis hispanica. The 2 methods are of similar sensitivity. The TMB-AHM reaction can be carried out at physiological pH (from 6-8), thus, avoiding tissue contraction. The reaction product is of an intense blue-green color and, as with the TMB-SNF method, shows granulation. The appearance of non-specific precipitates is completely avoided when the incubation medium is maintained at a pH 5.