Manual immunohistochemistry staining of mouse tissues using the avidin-biotin complex (ABC) technique.
Manual immunohistochemistry staining of mouse tissues using the avidin-biotin complex (ABC) technique.
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DOI：
10.1101/pdb.prot073429
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发表时间：
2014-06-02
影响因子：
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通讯作者：
Munn, Robert J
中科院分区：
文献类型：
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作者：
Cardiff, Robert D;Miller, Claramae H;Munn, Robert J
免疫组织化学 (IHC) 程序有许多变化，但都是基于将一抗附着到细胞上或细胞内的独特表位。此步骤之后是将细胞/一抗复合物与另一种二抗一起孵育，该二抗可识别产生一抗的物种。二抗上附着有指示分子。该指示剂在原始表位位点产生有色反应产物，从而实现可视化。这种基本的双抗体“三明治”程序有许多修改，包括其他层的抗体和许多指示剂，但所有变化都取决于抗体识别特定表位或抗原决定簇的独特能力。这里描述的程序称为 ABC（亲和素-生物素复合物）技术。该方法利用生物素对亲和素的高亲合力，从而形成牢固的键。该技术中描述的试剂产生金/棕色反应产物，可识别感兴趣的表位。
There are many variations on the immunohistochemistry (IHC) procedure, but all are based on attachment of a primary antibody to a unique epitope on or within the cell. This step is followed by incubation of the cell/primary antibody complex with another, secondary antibody that recognizes the species in which the primary antibody was produced. The secondary antibody has an indicator molecule attached to it. The indicator produces a colored reaction product at the site of original epitope, allowing visualization. This basic two-antibody "sandwich" procedure has many modifications that include other layers of antibodies and numerous indicators, but all variations depend upon the unique ability of antibodies to recognize specific epitopes or antigenic determinants. The procedure described here is called the ABC (avidin-biotin complex) technique. The method utilizes the high avidity of biotin for avidin, which allows formation of a strong bond. The reagents described in this technique produce a gold/brown reaction product that identifies the epitope of interest.