In this video, we give you tips for optimizing your Immunohistochemistry (IHC) techniques based on our experience at Cell Signaling Technology (CST).
Immunohistochemistry (IHC) is a powerful microscope-based technique that uses an antibody to view a specific protein in biological tissue. However, there are lots of steps in the procedure that each add variation that can impact final results. Therefore, individuals new to the method can struggle initially. All CST antibodies are developed and validated in-house so if the antibody you’re interested has been validated for IHC for paraffin-embedded samples (IHCP) or frozen samples (IHCF), the product’s Data Sheet will have our optimized assay conditions. However, if you need to deviate from our recommendations for some reason this video gives you tips and suggestions on re-optimizing so you can get reliable, reproducible results.
First, we’ll focus on how the different steps of the IHC protocol, like using either citrate or EDTA during the antigen retrieval step, impacts final staining. We’ll also go over ways to validate the specificity of your signal, like using treated paraffin-embedded cell pellets as positive and negative control, so you can trust the accuracy of your results. Finally, we give you examples on how the reagents you use can really make a difference on the strength of your final signal.
For more IHC tips, visit www.cellsignal.com bit.ly/1JhEn5D