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Immunohistochemistry

Overview
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Immunohistochemistry (IHC) is a powerful tool that uses the ability of specific antibodies to detect antigens of interest in tissue samples. IHC is widely used in the pathological evaluation of cancer tissues, to predict the prognosis of tumors, to predict responses to therapy, to confirm presence of infections, and in numerous research applications.1 In addition to detecting the presence or absence of an antigen, IHC also allows the determination of antigen distribution and location.

 

IHC Protocols

 

 

Principles of immunohistochemistry

The tissue of interest is first fixed, paraffin embedded or cryopreserved, and sectioned. The stored sections are processed (using deparaffinization for paraffin-embedded sections or by heating for cryopreserved sections) for antigen retrieval and then immunostained using antibodies specific to the antigens. Just like western blotting or ELISA, IHC can be direct or indirect (using only one antibody or a combination of a primary and a secondary antibody). Detection is usually accomplished by using enzymes such as horseradish peroxidase (HRP) or alkaline phosphatase (AP). Samples are visualized using microscopy.

 

Immunocytochemistry, immunohistochemistry and immunofluorescence—what are the differences?

The principles of immunohistochemistry are similar to those of immunocytochemistry (ICC), but IHC uses tissue samples while ICC uses cells. 

 

Both immunocytochemistry (ICC) and immunohistochemistry (IHC) employ immune labeling of antigens for analysis and use immunofluorescence methods. Sometimes, immunocytochemistry and immunofluorescence are used interchangeably. 

 

While ICC focuses on analysis at a cellular level, IHC allows examination of the whole tissue. Both techniques use immunofluorescence for detection. It is also possible to use enzyme-based detection (for example, using 3,3’-diaminobenzidine (DAB)).

References

  1. Duraiyan J, Govindarajan R, Kaliyappan K, Palanisamy M. Applications of immunohistochemistry. J Pharm Bioallied Sci. 2012;4(Suppl 2):S307-S309. doi: 10.4103/0975-7406.100281

IHC Sample Preparation
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IHC Detection Methods
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