Development of a highly sensitive immunohistochemical method to detect neurochemical molecules in formalin-fixed and paraffin-embedded tissues from autopsied human brains.
Development of a highly sensitive immunohistochemical method to detect neurochemical molecules in formalin-fixed and paraffin-embedded tissues from autopsied human brains.
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DOI：
10.3389/fnana.2015.00022
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发表时间：
2015
影响因子：
2.9
通讯作者：
Kaji R
中科院分区：
文献类型：
--
作者：
Goto S;Morigaki R;Okita S;Nagahiro S;Kaji R
免疫组织化学（IHC）是通过靶抗原与用可见标记物（例如，过氧化物酶）。我们已经开发了一种免疫染色方法，是高度敏感的神经化学抗原的检测。我们的IHC方法，我们称之为PBTA方法，涉及一种混合方案，该方案结合生物素-酪胺扩增实现聚合物和抗生物素蛋白-生物素复合物（ABC）方法的各个方面。当使用[Met]-脑啡肽作为靶抗原时，PBTA方法用于IHC的灵敏度比聚合物和ABC方法高100倍以上。此外，其酶联免疫吸附测定的灵敏度比ABC法高约1,000倍。我们研究了我们的IHC方法的实用性，用于显色和荧光检测系统，用于可视化神经化学肽和蛋白质在福尔马林固定，石蜡包埋组织从尸检人脑。结果令人信服地证明，在最佳条件下，我们的IHC方法是高度灵敏的，而不会增加非特异性背景活动。我们的免疫组化方法可能是一个强大的工具，用于检测和可视化的神经化学抗原，即使是在尸检人脑中的痕量。
Immunohistochemistry (IHC) is a valuable method for identifying discrete neurochemical molecules by the interaction of target antigens with validated antibodies tagged with a visible label (e.g., peroxidase). We have developed an immunostaining method that is highly sensitive in detection of neurochemical antigens. Our IHC method, which we call the PBTA method, involves a hybrid protocol that implements aspects of both the polymer and avidin-biotin-complex (ABC) methods in combination with biotin-tyramide amplification. When using [Met]-enkephalin as a target antigen, the sensitivity of the PBTA method for IHC was more than 100-fold higher compared with the polymer and ABC methods. In addition, its sensitivity for enzyme-linked immunosorbent assay was about 1,000-fold higher compared with the ABC method. We examined the utility of our IHC method for both chromogenic and fluorescence detection systems used to visualize neurochemical peptides and proteins in formalin-fixed, paraffin-embedded tissues from autopsied human brains. The results convincingly demonstrate that under optimal conditions, our IHC method is highly sensitive without increasing non-specific background activities. Our IHC method could be a powerful tool for detection and visualization of neurochemical antigens that are present even in trace amounts in autopsied human brains.
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影响因子：
5.3
作者：
Koizumi H;Morigaki R;Okita S;Nagahiro S;Kaji R;Nakagawa M;Goto S
通讯作者：
Goto S
影响因子：
2.9
作者：
Crittenden JR;Graybiel AM
通讯作者：
Graybiel AM
影响因子：
2.5
作者：
ROSSI, F;VANDERWANT, JJL;STRATA, P
通讯作者：
STRATA, P
影响因子：
3.2
作者：
WOOD, GS;WARNKE, R
通讯作者：
WARNKE, R
影响因子：
3.2
作者：
Hunyady, B;Krempels, K;Mezey, E
通讯作者：
Mezey, E