What Are the Key Steps in Quantitative Immunohistochemistry?
There are several crucial steps involved in qIHC:
Sample Preparation: Tissue samples are fixed, usually with formaldehyde, and embedded in paraffin. Sections are then cut and placed on slides. Antigen Retrieval: This step involves treating the tissue sections to unmask the epitopes, making them accessible to antibodies. Blocking: Non-specific binding sites are blocked to reduce background noise. Primary Antibody Incubation: Sections are incubated with a primary antibody that binds to the protein of interest. Secondary Antibody Incubation: A secondary antibody, conjugated with a detectable marker, is applied to bind the primary antibody. Detection: The signal from the marker is detected and quantified.
