Immunohistochemistry (IHC) Handbook

Immunohistochemistry (IHC) and immunocytochemistry (ICC) are techniques that use antibodies to detect antigens and provide semi-quantitative data about target protein expression, distribution, and localization. Both IHC and ICC are dependent on specific epitope-antibody interactions, but IHC refers to the use of tissue sections whereas ICC is performed on cultured cells or a cell suspension. In both of these methods, positive marker staining is visualized via a molecular label, which can be chromogenic (enzymatic) or fluorescent. The most challenging aspect of IHC is determining and optimizing the experimental conditions required to generate a strong and specific signal for each antigen of interest. While the technique is relatively straightforward in principle, there are many variables to consider in a given experiment.

For example, visualization of an abundant protein in formalin-fixed tissue will likely require antigen retrieval and may be compatible with direct detection using a fluorochrome-conjugated primary antibody. In contrast, detection of a phosphorylation-dependent epitope in a section of frozen tissue may require signal amplification and additional blocking steps.

In this handbook you'll learn more about IHC workflow, visual workflow, troubleshooting, and basic buffer and reagent recipes.