DOI: 10.3791/52293-v
This introductory level protocol describes the reagents, equipment, and techniques required to complete immunohistochemical staining of rodent brains, using markers for microglia and neuronal elements as an example.
The overall goal of the following experiment is to use immunohistochemistry in order to visualize proteins of interest. For this example, IBA one for microglial and pan neuronal for neurons. This is achieved by first placing prepared sections into a blocking solution to prevent non-specific binding.
As a second step antibodies, which recognize the two different proteins of interest are placed on the sections overnight. Next sections are washed and incubated in secondary antibodies in order to add fluorescent tags to the primary antibodies. The dual labeling results can point out microglial stainings within fields of neurons.
