FACTORS AFFECTING THE CYTOCHEMICAL REACTION OF PEROXIDASE WITH BENZIDINE AND THE STABILITY OF THE BLUE REACTION PRODUCT
FACTORS AFFECTING THE CYTOCHEMICAL REACTION OF PEROXIDASE WITH BENZIDINE AND THE STABILITY OF THE BLUE REACTION PRODUCT
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影响过氧化物酶与联苯胺细胞化学反应的因素及蓝色反应产物的稳定性
DOI：
10.1177/12.6.462
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发表时间：
1964
影响因子：
3.2
通讯作者：
W. Straus
中科院分区：
文献类型：
--
作者：
W. Straus
考察了pH、温度、培养时间、底物和受体的浓度、醇和盐的加入对过氧化物酶与联苯胺和H2O2细胞化学反应的影响。考察了蓝、棕反应产物对氧化还原剂的稳定性。结果表明，微酸性pH和低温是过氧化物酶与联苯胺反应产物生成蓝色而非棕色或黄色的主要因素。描述了一种稳定蓝色反应产物的方法，该反应产物包括用70%的硝普钠酒精溶液处理组织，在pH 5.0下缓冲，然后用联苯胺染色。通过调节过氧化物酶产生的“联苯胺蓝”的量，可以在没有添加盐（缓冲液）的情况下防止蓝色色素的结晶伪影。调节的主要变量是联苯胺和H2O2在0-4℃的孵育时间，次要变量是联苯胺、H2O2和添加醇的浓度。介绍了过氧化物酶在含醇和联苯胺水溶液中的染色方法，并对其进行了比较。讨论了过氧化物酶与联苯胺细胞化学反应的特异性。
The influence of pH, temperature, time of incubation, concentration of the substrate and acceptor, addition of alcohol and salts on the cytochemical reaction of peroxidase with benzidine and H2O2 was investigated. The stability of the blue and brown reaction products towards oxidizing and reducing agents was tested. It was observed that a slightly acid pH and low temperature are main factors for the formation of a blue rather than of brown or yellow reaction products of peroxidase with benzidine. A procedure was described for the stabilization of the blue reaction product consisting in the treatment of the tissue with 70% alcohol solutions of sodium nitroprusside, buffered at pH 5.0, subsequent to the staining with benzidine. Crystallization artifacts of the blue pigment could be prevented in the absence of added salts (buffers) by regulating the amount of "benzidine blue" produced by peroxidase. The main variables used for this regulation were the time of incubation with benzidine and H2O2 at 0-4°C, and secondary variables were the concentrations of benzidine, H2O2, and added alcohol. Procedures for the staining of peroxidase in alcohol-containing and in aqueous solutions of benzidine were described and compared. The specificity of the cytochemical reaction of peroxidase with benzidine was discussed.