Abstract
Immunohistochemistry (IHC) is a technique used to analyze protein expression in the context of tissue morphology. It exploits the specific binding of an antibody to its target epitopein combination with a chromogenic or fluorescent readout. Cell Signaling Technology (CST) antibodies validated for IHC have been developed in-house and CST scientists have determined the optimal conditions for the use of each antibody in IHC. The IHC protocol optimized for paraffin-embedded samples is detailed in this article so that you can replicate the procedure and obtain reproducible and reliable results. The main steps of this protocol are reagent and sample preparation, deparaffinization/rehydration, antigen unmasking (also called epitope or antigen retrieval), staining, dehydration, and coverslip mounting. Since all CST™ antibodies have been validated in-house using specific protocols, we strongly recommend following the protocol provided with each antibody. Some common areas where troubleshooting may be required are also discussed.
