Introduction to Immunohistochemistry (IHC)

The method of immunohistochemistry (IHC) demonstrates the distribution and localization of antigens, such as proteins, in tissue sections using the interaction between antibody and antigen.​​

The method is particularly useful for diagnosing abnormal cells in diseases like cancer. In general, the information gained from IHC, with its unique perspective, creates a context for data obtained using other methods.

Antibodies can recognize the target antigen and that is what enables the immunohistochemical staining. Since antibodies are highly specific, the antibody will bind only to the antigen of interest in the tissue section.

The antibody-antigen interaction is then visualized using either chromogenic detection, in which an enzyme conjugated to the antibody catalyzes the conversion of a substrate to produce a colored precipitate at the location of the antigen.

The other way to visualize the interaction is through a fluorescent detection, in which a fluorophore is conjugated to the antibody and can be visualized using fluorescence microscopy.

Materials and Reagents

  • Xylene
  • 100% ethanol
  • 95% ethanol
  • 70% ethanol
  • 50% ethanol

Method

Place the slides in a rack, and perform the following washes using a Coplins jar:

Solution Incubation time
Xylene 3 min
Xylene 3 min
1:1 Xylene:100% Ethanol 3 min
100% Ethanol 3 min
100% Ethanol 3 min
95% Ethanol 3 min
70% Ethanol 3 min
50% Ethanol 3 min

About Abcam

Abcam is a global life sciences company providing highly validated antibodies and other binders and assays to the research and clinical communities to help advance the understanding of biology and causes of disease.

Abcam’s mission is to serve life scientists to help them achieve their mission faster by listening to their needs, continuously innovating and improving and by giving them the tools, data and experience they want. Abcam’s ambition is to become the most influential life science company for researchers worldwide.


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Last updated: Jun 6, 2019 at 10:23 AM

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