Immunohistochemistry

Immunohistochemisty (IHC) can be used to determine the presence of a particular protein within a tissue, the distribution in that tissue, and its precise location within a single cell. There are many IHC methodologies available. It is important that the methodology used be sensitive, produce little or no background, and give reliable and reproducible results. IHC is used at the light microscope level. It is an enzyme-based detection method that includes the use of horseradish peroxidase (HRP) or alkaline phosphatase (AP). Key points to keep in mind in IHC include choosing an enzyme system, preparing tissue properly, and using the correct tissue staining techniques. The two most commonly used enzyme detection systems are the horseradish reoxidase and the alkaline phosphatase systems. The HRP or peroxidase systems are most widely used and tend to produce a dense label. The HRP system provides better staining of nerve axons and projections in the CNS. The AP or alkaline phosphatase systems are more sensitive and allow for better cell morphology. The AP system is also used when endogenous peroxidase activity is a problem.16

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