What are the Steps in an Immunohistochemistry Procedure?
The basic steps in an IHC procedure include:
Tissue Preparation: Tissues are fixed, usually in formalin, and embedded in paraffin. Thin sections are then cut and mounted on slides. Antigen Retrieval: Treatment of tissue sections to unmask antigens that may be hidden due to the fixation process. Blocking: Blocking non-specific binding sites to prevent background staining. Primary Antibody Incubation: Application of the primary antibody specific to the target antigen. Secondary Antibody Incubation: Application of a secondary antibody that recognizes the primary antibody and is conjugated to a detection enzyme or fluorophore. Detection: Visualization of the antigen-antibody complex using a chromogenic substrate (for IHC) or fluorescence (for IF).
