Immunohistochemical and Immunofluorescence Procedures for Protein Analysis
Immunohistochemical and Immunofluorescence Procedures for Protein Analysis
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DOI：
10.1007/978-1-61779-289-2_11
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发表时间：
2011-01-01
期刊：
影响因子：
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通讯作者：
O'Sullivan, Finbarr
中科院分区：
文献类型：
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作者：
Katikireddy, Kishore Reddy;O'Sullivan, Finbarr
免疫组织化学(IHC)和免疫荧光(IF)包括抗体与感兴趣的细胞或组织抗原结合，然后用荧光/3，3‘-二氨基联苯胺(DAB)显色系统显示结合产物。随着针对细胞表位的抗体数量的增加，IHC和IF是非常有用的诊断工具，也是指导针对细胞/组织样本上特定抗原的特定治疗的手段。根据研究样本的类型、所需的敏感性程度和成本考虑，可以使用几种IHC和IF染色方法。以下是通过直接、间接、IHC和IF定位蛋白质和其他抗原的基本“通用”方法。该方法依赖于组织/细胞的适当固定，以保持抗原的细胞分布和保持细胞的形态。概述了所需试剂的详细情况。还考虑了人工制品和其他潜在的陷阱，从而采取了避免它们的手段。
Immunohistochemistry (IHC) and immunofluorescence (IF) involve the binding of an antibody to a cellular or tissue antigen of interest and then visualisation of the bound product by fluorescence/with the 3,3'-diaminobenzidine (DAB) chromogen detection system. With increasing numbers of available antibodies against cellular epitopes, IHC and IF are very useful diagnostic tools as well as a means to guide specific therapies that target a particular antigen on cell/tissue samples.There are several IHC and IF staining methods that can be employed depending on the type of specimen under study, the degree of sensitivity required, and the cost considerations. The following is a basic "generic" method for localising proteins and other antigens by direct, indirect, IHC and IF. The method relies on proper fixation of tissue/cells to retain cellular distribution of antigen and to preserve cellular morphology. Details of reagents required are outlined. Consideration is also given to artefacts and other potential pitfalls and thus means to avoid them.