Sample Preparation: Tissue samples are collected, fixed in formalin, and embedded in paraffin. Thin sections are then cut and mounted on slides. Antigen Retrieval: To unmask antigens, slides undergo treatment with heat or enzymatic digestion. Blocking: Non-specific binding sites are blocked to prevent background staining. Primary Antibody Incubation: The primary antibody, which is specific to the target antigen, is applied. Secondary Antibody Incubation: A secondary antibody, conjugated with a detection enzyme or fluorophore, binds to the primary antibody. Detection: The enzyme or fluorophore generates a colorimetric or fluorescent signal, highlighting the presence of the target antigen. Counterstaining: Tissue sections are often counterstained to provide contrast.
