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Keywords: immunohistochemistry controls, IHC experiment design, control types
In the lab, researchers invest time and effort to optimize the sample preparation and sample staining processes of IHC. When successful, the results produce a strong and specific signal. However, how do we know the results have been interpreted correctly? When designing your IHC experiment, it is crucial to include positive and negative controls. The controls will help verify whether or not the IHC staining results observed are accurate and valid while exposing experimental artefacts. Read on to find out the 6 established controls for IHC!
Keywords: IHC optimization tips, antigen retrieval principle explanation, immunohistochemistry protocol optimize, how to perform HIER antigen retrieval for IHC experiment
If you are performing IHC experiments, you may need to optimize your antigen retrieval methods. Techniques generally fall into two main categories: protease-induced epitope retrieval (PIER) and heat-induced epitope retrieval (HIER). In general, HIER has a much higher success rate and is recommended over PIER methods. The protocol for HIER must be optimized for each sample tissue, fixation method, and antigen. This method is especially time-, temperature-, buffer-, and pH-sensitive, and the best condition must be determined empirically. Here is a step-wise protocol that may be helpful for your immunohistochemistry experiment.
Keywords: IHC, immunohistochemistry, heat induced epitope retrieval, proteolytic induced epitope retrieval, PIER, HIER, antigen retrieval optimization
Antigen retrieval is often necessary when performing IHC experiments. Here are some key points to know about HIER and PIER methods of antigen retrieval. This protocol step needs to be optimized for every antigen of interest.
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