The IHC process typically involves several key steps:
1. Tissue Preparation: Fixation and embedding of tissues, usually in paraffin, followed by sectioning. 2. Antigen Retrieval: Treatment of tissue sections to unmask antigens, often using heat-induced or enzyme-induced methods. 3. Blocking: Application of blocking reagents to reduce non-specific binding. 4. Primary Antibody Incubation: Application of the primary antibody to bind the target antigen. 5. Secondary Antibody Incubation: Application of the secondary antibody to bind the primary antibody. 6. Detection: Application of chromogenic substrates to visualize the bound antibodies. 7. Counterstaining: Application of counterstains to provide contrast. 8. Mounting: Covering the stained tissue section with a coverslip for viewing under a microscope.
