title: Immunohistochemical and immunofluorescence procedures for protein analysis.
abstract: 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.
authors: National Institute of Cellular Biotechnology, Dublin City University, Ireland. kishore.katikireddy2@mail.dcu.ie
authors: Kishore Reddy
authors: KR
authors: Katikireddy
authors: Finbarr
authors: F
authors: O'Sullivan
mesh_terms: D015100:3,3'-Diaminobenzidine / Q000032:analysis / Q000737:chemistry; D000941:Antigens / Q000032:analysis*; D005404:Fixatives / Q000737:chemistry; D005455:Fluorescent Antibody Technique / Q000379:methods*; D020869:Gene Expression Profiling; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D016612:Paraffin Embedding; D011506:Proteins / Q000032:analysis* / Q000235:genetics; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods
chemical_list: D000941:Antigens; D005404:Fixatives; D011506:Proteins; D015100:3,3'-Diaminobenzidine