Immunohistochemistry / Immunofluorescence
I. Required material
Reagents
- A primary antibody , labeled or not, against the target molecule
- Revelation kit
- Chromogen
Buffers
- Xylene
- Alcooholic baths at 70%, 80%and 95%
- Hydrogen peroxide (H2O2)
- Tris EDTA, Tris HCl
- Tween-20
Material
- Pipets and Pipet-aid
- Water bath
II. Experimentation length
- 10 mn of peroxydase blocking
- 30 mn of antigenic retrieval
- 1 h incubation of primary antibody
- 10 mn incubation...
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The principle of ABC detection kits is based on the affinity of avidin (or streptavidin) for biotin. Indeed, abbreviation ABC stands for Avidin / Biotin Complex.
In general, the ABC detection kits are composed of:
- A biotinylated secondary antibody
- An Avidin / Enzyme Complex
The kits may or may not contain the blocking and dilution buffers and the substrate depending on the formats.
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