The process of single marker IHC involves several steps: 1. Tissue Preparation: Tissue samples are fixed and embedded in paraffin to preserve morphology. 2. Antigen Retrieval: Techniques like heat-induced epitope retrieval (HIER) are used to unmask antigens that may be hidden due to fixation. 3. Blocking: Non-specific binding sites are blocked to prevent background staining. 4. Primary Antibody Application: A primary antibody specific to the antigen of interest is applied to the tissue section. 5. Secondary Antibody Application: A secondary antibody, conjugated with a detection label (such as an enzyme or fluorophore), binds to the primary antibody. 6. Visualization: The bound antibodies are visualized using appropriate substrates or detection systems.
