This whitepaper is a complete guide to inmunohistochemistry, exploring in great detail the state of the art in this field. This guide provides a starting point for those researchers who are taking their first steps on inmunohistochemistry, and at the same time acts as a chance for the more veteran ones to keep up to date with the latest developments on the field.
Immunohistochemistry (IHC) and immunocytochemistry (ICC) are powerful techniques that use the antibody-antigen affinity to detect antigens (proteins). Antibody-based applications rely on the specific binding of antibody to the target epitope to generate accurate expression data. Antibodies specifically recognize the epitope of interest through intermolecular forces including hydrophobic interactions, ionic interactions, hydrogen bonding, and others.
Both techniques, provide specific semi-quantitative data about target protein expression, localization, and distribution . Thus, the observation of processes in the context of intact tissue is possible using any of those techniques, even though they are less sensitive quantitatively than other immunoassays, such as ELISA.
This guide contains:
- What does inmunohistochemistry mean?
- Experimental desing
- Sample prep and sample fixation
- Antigen retrieval
- Blocking and Detection
- Microscopy
- Quantitative image processing
- Controls
- Troubleshooting
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Choosing an antigen: Peptide or protein
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