Encyclopedia of Experiments: Immunology
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Transcript
Begin with a slide carrying a formalin-fixed, paraffin-embedded rabies virus-infected mouse brain tissue section. The fixed section retains viral antigens via protein cross-linking.
Treat the slide with a xylene solution to remove the paraffin.
Immerse the slide in decreasing ethanol concentrations to rehydrate the tissue, making it compatible with aqueous solutions.
Introduce a protease-containing buffer to disrupt protein cross-links, improving antigen accessibility.
Submerge in hydrogen peroxide to block endogenous peroxidases, and apply a blocking solution to prevent non-specific interactions.
Introduce primary antibodies specific to the rabies antigen, forming antigen-antibody complexes.
Add biotinylated secondary antibodies, which bind to the primary antibodies. Incubate with streptavidin-conjugated peroxidase enzymes, which interact with biotin.
Introduce a peroxidase substrate, producing a magenta-red precipitate.
Counterstain the tissue with a nuclear stain, followed by a bluing agent, staining the nuclei blue.
The immunohistochemistry image reveals magenta-red spots in the cytoplasm, confirming the presence of rabies antigens in the tissue section.
