Mycoplasma Protocols

Volume 104 of the series Methods in Molecular Biology™ pp 133-140

Immunohistochemical Staining of Fixed Tissues

  • Eugenio ScanzianiAffiliated withIstituto di Anatomia Patologica Veterinaria e Patologica Aviare

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Immunohistochemistry is a technique in which the specific interaction between an immunoglobulin and its homologous antigen is visualized on histological sections by a microscopically detectable label. Generally, the label consists of an enzyme, such as peroxidase, alkaline phosphatase, or glucose oxidase that reacts with an appropriate substrate-chromogen solution to produce a specific color at the site of reaction. Several immunohistochemical techniques have been developed and the most important are schematically represented in Fig. 1. In the direct method, the primary antibody is directly labeled with the enzyme. In the indirect method, an enzyme-labeled secondary antibody is directed against the immunoglobulin type of the animal species in which the primary antibody has been raised. Both methods have a relatively low sensitivity and are therefore not frequently used.
Fig. 1.

Schematic representation of immunohistochemical techniques. Direct method, indirect method, ([A] left to right) PAP complex procedure, ABC procedure ([B] left to right). A=avidin; B=biotin; C=substrate-chromogen; P=peroxidase.