Immunohistochemistry (IHC) Methods, Techniques & Protocols

Definition: localization of antigens or proteins in tissue sections by the use of labeled antibodies as specific reagents through antigen-antibody interactions that are visualized by a marker such as fluorescent dye, enzyme, or colloidal gold.

Introduction to Immunohistochemistry (IHC World) - Immunohistochemistry is the localization of antigens in tissue sections by the use of labeled antibodies as specific reagents through antigen-antibody interactions that are visualized by a marker such as fluorescent dye, enzyme, radioactive element or colloidal gold.

Immunohistochemistry Protocol Database (IHC World) - This protocol database is compiled based on published literatures, laboratory submissions, as well as individual experimental data. It contains antibody staining protocols, antigen retrieval protocols, and more.

Antibody Staining Protocols (IHC World) - A comprehensive list of antibody staining protocols for immunohistochemistry application, including detailed procedure, technical notes, washing buffer recipes, etc.

Antigen Retrieval Protocols (IHC World) - The most complete list of antigen retrieval protocols, including, Heat Induced Epitope Retrieval (HIER), Proteolytic Induced Epitope Retrieval (PIER), Frozen Section Epitope Retrieval, Universal Antigen Retrieval Method, Free Floating Section Epitope Retrieval.

General Immunohistochemistry Protocols (IHC World) - Standarded immunohistochemistry protocols, double labeling protocols, immunoenzyme methods, immunofluorescence methods.

Immunohistochemistry Techniques (NordiQC) - Immunohistochemistry is technically complex, and no aspect of this complexity can be ignored, from the moment of collecting the specimen to issuance of the final report (Taylor CR. Arch Pathol Lab Med 2000; 124:945). The techniques pages give you some hint regarding optimizing the various steps in your protocol. The 'Controls' page illustrate a method of producing multi-tissue blocks.

Recommended IHC protocols from NordiQC assessment schemes - Among protocols shown to give optimal staining results, one or more are selected to cover a spectrum of laboratories, antibodies and techniques. Only the latest recommended protocols for each antibody/clone/epitope are listed here.

IHC Staining Protocol for Mouse Antibody On Mouse Tissue (IHC World) - Antigen detection with mouse primary antibody on mouse tissues is complicated by high levels of background staining when indirect immunohistochemical detection methods are used. The main cause of the background staining is due to the binding of secondary anti-mouse antibody to endogenous mouse tissue Igs and other components. Blocking this binding by pre-incubation with Fab Fragment of Unconjugated Anti-Mouse IgG in combination with the use of a biotin-conjugated Fab Fragment Anti-Mouse IgG (in place of whole labeled secondary antibody) led to the most complete elimination of background staining and achieved satisfactory result. This blocking method can be also adapted to the use of other antibodies on homologous tissues.

Immunohistochemistry on Whole-Mount Embryos (Paul MacDonald)