A stable and highly sensitive 3,3′,5,5′-tetramethylbenzidine-based substrate reagent for enzyme-linked immunosorbent assays
A stable and highly sensitive 3,3′,5,5′-tetramethylbenzidine-based substrate reagent for enzyme-linked immunosorbent assays
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一种基于 3,3′,5,5′-四甲基联苯胺的稳定且高灵敏度的底物试剂，用于酶联免疫吸附试验
DOI：
10.1016/s0022-1759(99)00166-0
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发表时间：
2000-01-13
影响因子：
2.2
通讯作者：
Schmidt, MA
中科院分区：
文献类型：
--
作者：
Frey, A;Meckelein, B;Schmidt, MA
3，3‘，5，5’-四甲基联苯胺(TMB)是一种广泛应用于辣根过氧化物酶检测系统的显色剂，其反应产物吸收系数高，无致癌性。不幸的是，TMB不稳定，在水溶液中不易溶解，必须在每次实验前新鲜制备此类溶液。此外，在分析条件下可能会发生底物耗尽。为了克服这些问题，我们开发了一种双组分TMB底物系统，与许多商用TMB试剂相比，该系统具有更低的检测下限和更高的灵敏度。底物体系的两个组分在4摄氏度下至少稳定一年，通过添加稳定剂防止TMB原液通常变色，稳定剂在两个组分混合时分解。(C)2000 Elsevier Science B.V.保留所有权利。
3,3',5,5'-Tetramethylbenzidine (TMB) is a widely used chromogen for horseradish peroxidase-based detection systems because it yields reaction products with high absorption coefficients and lacks carcinogenicity. Unfortunately, TMB is labile and poorly soluble in aqueous buffers and such solutions must be freshly prepared before each experiment. Moreover, substrate depletion can occur under assay conditions. To overcome these problems we have developed a two-component TMB substrate system which has a lower detection limit and is more sensitive than many commercially available TMB reagents when compared in microtiter plate enzyme-linked immunosorbent assays. Both components of the substrate system are stable for at least 1 year at 4 degrees C and the usual discoloration of TMB stock solutions is prevented by the addition of a stabilizer that decomposes upon mixing of the two components. (C) 2000 Elsevier Science B.V. All rights reserved.