UNIT 14.6 Immunohistochemistry

This unit surveys immunohistochemical techniques for protein localization. The first four protocols focus on indirect immunofluorescence which is the easiest and most widely used method of optical detection. Additional protocols present methods suitable for bright-field microscopy, such as immunoperoxidase and immunogold labeling. The commentary contains an extensive troubleshooting guide with a list of suitable routine controls. The chosen method of tissue or cellular preparation must be suitable to answer the fundamental objective of the protocol (i.e., where the protein is localized within the system). Furthermore, the choice of fluorochrome and the method of labeling are also important to optimize the sensitivity of antigen localization. The basic protocol is therefore divided into two sections: (1) cellular preparations (monolayers of adherent cells and cells grown in suspension) and (2) tissue section labeling.