Immunohistochemistry Protocol for Paraffin embedded Tissue Sections

IHC Protocol Video for Paraffin-embedded Tissue Sections from Cell Signaling Technology (CST)
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Immunohistochemistry (IHC) is a powerful microscope-based technique that uses an antibody to view a specific protein in biological tissue. It is often used to diagnose abnormal cells in solid tumors, visualize molecular markers for cellular events like apoptosis, and monitor the localizations and expression of biomarkers. However, individuals new to this method can struggle due to the many steps in the procedure that can each add variations that will impact staining.

Cell Signaling Technology (CST) scientists have determined optimal conditions for CST antibodies developed and validated in-house for IHC. In this video, we demonstrate the IHC protocol we have optimized for formalin-fixed paraffin embedded (FFPE) tissue sections so you can replicate results in your laboratory and obtain consistent results.

We will first describe how to prepare your sample and deparaffinization/rehydrate the FFPE samples followed by the antigen unmasking step, the most difficult step in an IHC protocol. Next, we will review chromogenic staining with the SignalStain DAB Substrate Kit followed by how to mount sections to a coverslip before viewing on a microscope. Finally, we describe how using different conditions than those recommended each antibody Product Data Sheet, like changing the recommended antibody diluent, can affect your final results.

If you’re interested alternate protocols including performing IHC on frozen tissue samples visit the Protocol at the Cell Signaling Technology website (http://cellsignal.com/protocols).