Immunohistochemistry (IHC) is an antibody-based technique that detects antigens of interest in fixed tissue sections, often visualized by light microscopy (see figure below). In this multi-step application, staining is highly influenced by multiple variables. High-quality reagents and optimized protocols are key for generating accurate expression, localization and distribution data. Our complete line of IHC products and scientific resources will help you produce valid, reproducible results you can trust.
IHC Detection Methods. As illustrated above, enzymatic conversion of a chromogen substrate is required to visualize an antigen in traditional IHC experiments. Fluorescence detection is an alternative method that is more suitable for multicolor experiments due to the broad availability of fluorochromes and the emergence of high contrast imaging. Read more about IHC detection methods.
The most critical factor to achieve successful staining is selecting an antibody that specifically binds the target antigen.
In indirect detection, a labeled secondary antibody (fluorescent, enzymatic, biotin, etc.) and, in some cases, a third layer for signal amplification is required to detect the antigen-primary antibody complex. Compared to directly labeled primary antibodies, indirect detection is more sensitive and vital for effective identification of low abundance antigens, rare epitopes, and under conditions of nonoptimal antigen-antibody binding. Read more about detection methods.