Immunohistochemistry / Immunofluorescence
Immunohistochemistry (IHC) General protocol
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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Immunohistochemistry protocol - Avidin/Biotin Method (ABC)
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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