A novel immunohistochemical method for evaluation of antibody specificity and detection of labile targets in biological tissue
A novel immunohistochemical method for evaluation of antibody specificity and detection of labile targets in biological tissue
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DOI：
10.1016/s0165-022x(03)00149-0
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发表时间：
2004-01-30
影响因子：
--
通讯作者：
Levin, M
中科院分区：
文献类型：
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作者：
Levin, M
免疫组织化学是分析组织内靶分子定位的关键工具。然而，准确的结果需要一种能够区分相似化合物的抗体。我们提出了一种简单的免疫组织化学方法，也可用于快速评估抗体的特异性。我们用血清素（一种极不稳定的化合物）证明了这一技术。5-羟色胺（5-hydroxytryptamine, 5HT）是一种重要的神经递质，可调节正常认知和多种精神障碍，以及肿瘤生长、心脏病理和胚胎发生。5 -羟色胺的免疫组织化学检测通常被用作神经元细胞标志物，并提供5 -羟色胺作为胚胎形态形成因子的重要信息。有必要能够区分血清素与密切相关的分子具有显著不同的生物活性。使用我们的方法，我们鉴定了血清素特异性抗体，并表明在已发表的论文中经常用于鉴定血清素能细胞的一些商业5HT抗体不是5HT特异性的。这些数据证明了特异性检测抗体的必要性（特别是在高临床相关性的领域，如5HT）。我们也说明检测血清素能细胞在胚胎组织使用我们的技术。这种方法为测试抗体对任何生物分子的特异性提供了许多普遍的优点，并有助于避免在免疫组织化学过程中的假阳性和阴性。(C) 2003, Elsevier B.V.出版
Immunohistochemistry is a key tool for analyzing target molecule localization within tissues. However, accurate results require an antibody that can distinguish between similar compounds. We present a simple immunohistochemical method that can also be used to rapidly evaluate antibodies' specificities. We demonstrate this technique with serotonin, an extremely labile compound. Serotonin (5-hydroxytryptamine, 5HT) is an important neurotransmitter regulating normal cognition and several mental disorders, as well as tumor growth, cardiopathology, and embryogenesis. In-imunohistochemical detection of serotonin is commonly used as a neuronal cell marker and to provide crucial information on serotonin's role as an embryonic morphogen. It is necessary to be able to distinguish serotonin from closely related molecules with significantly different biological activity. Using our method, we identify antibodies that are specific for serotonin and show that some commercial 5HT antibodies often used to identify serotonergic cells in published papers are not 5HT-specific. These data demonstrate the necessity of specifically testing antibodies (especially in areas of high clinical relevance such as 5HT). We also illustrate detection of serotonergic cells in embryonic tissue using our technique. This method offers a number of general advantages for testing specificity of antibodies to any biological molecule, and helps avoid false positives and negatives during imimmohistochernistry. (C) 2003 Published by Elsevier B.V.