1. Primary Antibody Binding: A primary antibody specific to the target antigen is applied to the tissue section. 2. Secondary Antibody Conjugation: A secondary antibody conjugated with horseradish peroxidase (HRP) is then applied. This secondary antibody binds to the primary antibody. 3. Tyramide Activation: Tyramide conjugates (such as tyramide-fluorescein) are introduced. The HRP enzyme catalyzes the activation of the tyramide, creating highly reactive intermediates. 4. Signal Amplification: These intermediates covalently bind to nearby tyrosine residues on proteins within the tissue, leading to an amplified signal that can be detected using various methods like fluorescence or chromogenic substrates.
