Antigen retrieval

Detection techniques using antibodies often fail to work on PFA- or formalin-fixed, paraffin-embedded sections. Antigen retrieval methods can then, in some cases, enable specific antibody detection. They work by reversing some of the chemical modification of epitopes during fixation. These procedures will not help much with epitope loss due to denaturation during sample treatment, like hot paraffin embedding.

Heat treatment

 * Mechanism: heat cleaves cross-links, exposes buried epitopes of proteins, protein unfolding/refolding
 * Specific protocol 1: citrate buffer
 * Specific protocol 2: Tris-EDTA buffer

Protease treatment

 * Specific protocol 1: proteinase K
 * Specific protocol 2: trypsin

Detergent treatment

 * Specific protocol: SDS

Personal and lab-specific protocols

 * Korey Griffin's notes:
 * Griffin:Antigen Retrieval Technique - overview and comparison of techniques
 * Griffin:Immunohistochemistry_Paraffin - heat-based retrieval
 * Griffin:Immunofluorescence_Cell_Staining#Antigen_retrieval - detergent-based retrieval


 * Livesey's antigen retrival - heat-based retrieval
 * Immunohistochemistry for AntiOsteocalcin Antibody - a protocol with a protease-based antigen retrieval