Heat-Induced Epitope Retrieval
Sections are heated to roughly 95-120 C in a citrate (pH 6) or EDTA/Tris (pH 8-9) buffer, which hydrolyzes methylene cross-links and chelates the calcium complexes that hold masked conformations in place. Buffer pH, temperature and time form a three-variable window that must be optimized per antibody.
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Sections are heated to roughly 95-120 C in a citrate (pH 6) or EDTA/Tris (pH 8-9) buffer, which hydrolyzes methylene cross-links and chelates the calcium complexes that hold masked conformations in place. Buffer pH, temperature and time form a three-variable window that must be optimized per antibody.
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