ICC/IHC Protocols

Primary Antibody Selection & Optimization

The most important factor when designing an IHC/ICC experiment is selection of the primary antibody. In turn, the critical feature of a primary antibody is specificity for the epitope. All steps of an IHC/ICC experiment must be optimized to visualize specific staining and minimize non-specific background signals. This includes performing initial studies to determine the appropriate incubation conditions for each primary antibody.

Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections

In order for tissue sections to be retained on histological slides during staining and washing steps, slides need to be coated with adhesive compounds. Although there are a variety of such compounds, gelatin is the most frequently used for histological purposes.

Please read the protocol in its entirety before starting.

Protocol for Making a 4% Formaldehyde Solution in PBS

The vast majority of IHC/ICC procedures employ fixation of tissues and cells using formaldehyde-based fixatives. The protocol below describes the technique for generating a 4% formaldehyde solution in PBS. The most effective fixative must be determined experimentally.

Caution: Formaldehyde is toxic. Please read the MSDS before working with this chemical. Gloves and safety glasses should be worn and solutions made inside a fume hood.

Please read the protocol in its entirety before starting.