IHC Principle
Western blotting, ELISAs, and IHC are all immunoassays, and so all share in common the same basic principle of operation: primary antibodies bind to the target protein of interest, then labelled secondary antibodies bind the primary antibody, allowing for the precise detection of protein.
IHC is the application of the immunoassay principle to tissue staining. By using antibodies to direct staining, IHC can accurately detect protein localization while generating crisp stains of tissue structures.
IHC Sample Preparation
Every immunoassay protocol begins with sample preparation. Western blot, IHC, and ELISA all require different ways of preparing a sample, and nearly every sample type requires specialized treatment in order to produce high-quality, consistent results.
IHC, unlike Western blot or ELISA, does not require that proteins be extracted from cells or tissues. Instead, sample preparation for IHC focuses on properly fixing, slicing, and mounting the samples. Use these sample preparation guides to learn how to prepare cell cultures and various tissue types for IHC.
IHC Protocol
The Boster Bio protocol for IHC offers a step-by-step overview of the procedure. Use this guide as a primer or a quick reference guide, and see our product datasheets or sample preparation guides for more detailed information.
IHC Optimization Tips
Nearly every step of the IHC procedure comes with a choice. Which sample preparation method is best? How concentrated should the antibodies be? Which blocking buffer will work best?
The Boster Bio optimization tips are meant to help answer those questions and more. Take a look at our optimization guides to help inform your decisions, optimize your experiments, and get better results.
IHC Troubleshooting Tips
Every researcher eventually encounters trouble with an experiment. Blots appear blank, or IHC stains come out blurry, or ELISA standard curves come out wonky, or any of a thousand different problems.
Proper controls can help eliminate a great number of possible sources of error, but many troubles will remain. Check out the Boster troubleshooting guides for the most common troubles, their sources, and how to resolve them.