IHC for paraffin-embedded sections video protocol

This video demonstrates immunohistochemistry (IHC) with paraffin sections, using variety of antibodies and stains to reveal the molecular architecture of the tissue.

Paraffin-embedded sections are first dewaxed to replace the wax with water; this is because most staining solutions are aqueous.

Dewaxing is usually followed by an antigen-retrieval method, which unmasks antigens that have been obscured during the fixation process.

Next, depending upon the type of antibodies that will be used, a series of blocking steps take place that serve to minimize non-specific interactions of the antibody and block endogenous enzymes that can result in false positives.

After blocking, the sections are incubated in antibodies; this incubation can be in directly conjugated primaries or can consist of sequential incubations in both primary and secondary antibodies (fluorescent or enzyme-linked).

If an enzyme-linked secondary is used, a further incubation in substrate will be required, otherwise the sections can be mounted and imaged.

View our IHC staining protocol for paraffin, frozen and free floating sections.