Immunohistochemistry Protocols


Your source for protocols, techniques and information about immunohistochemistry, immunology, histology, pathology and chemistry.

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Immunohistochemistry Protocols

This site is dedicated to bringing basic as well as advanced information concerning immunohistochemistry for both the researcher, histotechnician, academic and surgical pathologist. We welcome any feedback and suggestions.

Immunohistochemistry Technique

-  Slices of fresh tissue and placed in either B5 fixative or 10% formalin and paraffin embedded

-   Paraffin sections are created with a microtome, subjected to antigen-retrieval methods and immunohistochemical techniques

Immunohistochemistry Advantages

-    Economical

-    Done routinely in most hospitals (basic markers)

Immunohistochemistry Disadvantages

-    False positives and false negatives may result based on fixation, cross-reactivity, antigen retrieval, sensitivity of antibodies used, interpretetation errors, etc.

History of Immunohistochemistry

Immunohistochemistry is a molecular technique that combines principles from both immunology and biochemistry techniques and principles to the study of histology and pathology by revealing molecules and patterns within cells and tissues.

The first to describe immunohistochemistry was Dr. Coons. The original immunohistochemistry method consisted of an antibody tagged with a fluorescent probe which was developed in rabbits, which was mixed with tissue sections and searched for, under a fluorescent microscope following a period of incubation. Since, numerous advancements and improvements have been done, to make the technique of immunohistochemistry fairly inexpensive and indispensable in both pathology departments and molecular laboratory benches worldwide.

Numerous different procedures are available, however the most commonly used are the peroxidase-antiperoxidase immune complex method and more so, the biotin-avidin immunoenzymatic technique.