Postcoupling, noncoupling, and fluorescence techniques for the demonstration of alkaline phosphatase.
Postcoupling, noncoupling, and fluorescence techniques for the demonstration of alkaline phosphatase.
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用于演示碱性磷酸酶的后偶联、非偶联和荧光技术。
DOI：
10.1093/jnci/24.5.1199
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发表时间：
1960
期刊：
影响因子：
--
通讯作者：
M. S. Burstone
中科院分区：
文献类型：
--
作者：
M. S. Burstone
发展和使用新的后耦合，非耦合，和荧光技术的演示碱性磷酸酶的描述。该技术利用新的底物，羟基萘甲酸的衍生物和复合胺。三种最有用的底物是5，6，7，8-β-四氢萘酚羧酸-β-萘酰胺磷酸盐（I）、2-羟基-3-萘甲酸-2-氨基蒽醌酰胺磷酸盐（II）和2-羟基-3-萘甲酸-氨基偶氮苯甲酰苯胺磷酸盐（III）。该技术主要用于石蜡包埋组织，可提供精确的定位。这些比较的最好的结果，获得了通过非线性耦合技术。在一个基板的情况下，蒽醌衍生物（II），postfixed冷冻切片给出有用的结果。从底物I释放的萘酚是一种荧光染料，是酶组织化学中一种新途径的基础，即通过荧光观察酶活性。该底物和另一种底物也释放出萘酚，其在光学放大下是可见的，而无需后耦合。此应用程序称为非耦合程序。对于大多数目的，推荐使用底物II或III的后偶联程序。孵育溶液是稳定的，并且随后的后偶联可以用多种重氮盐来完成。这些技术也适用于证明碱性磷酸酶以外的水解酶。
The development and use of new post-coupling, noncoupling, and fluorescence techniques for the demonstration of alkaline phosphatase are described. The techniques utilize new substrates, derivatives of hydroxynaphthoic acid, and complex amines. Three of the most useful substrates are 5,6,7,8-β-tetralol carboxylic acid-β-naphthylamide phosphate (I), 2-hydroxy-3-naphthoic acid-2-aminoanthraquinonylamide phosphate (II), and 2-hydroxy-3-naphthoic acid-aminoazobenzanilide phosphate (III). The techniques, which are designed primarily for use with paraffin-embedded tissues, give sharp localizations. These compare with the best results obtained by the simultaneous-coupling technique. In the case of one substrate, an anthraquinone derivative (II), postfixed frozen sections give useful results. The naphthol released from substrate I is a fluorochrome and is the basis of a new avenue of approach in enzyme histochemistry, namely, the visualization of enzyme activity by fluorescence. This and another substrate also liberate naphthols, which are visible under optical magnification without postcoupling. This application is referred to as anoncouplingprocedure. For most purposes, a postcoupling procedure utilizing substrates II or III is recommended. The incubating solutions are stable, and subsequent postcoupling may be accomplished with a large variety of diazonium salts. These techniques also lend themselves to the demonstration of hydrolytic enzymes other than alkaline phosphatase.