Antigen retrieval

From OpenWetWare
Jump to navigationJump to search
back to protocols

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.

Types of retrieval methods

Heat treatment

  • Mechanism: heat cleaves cross-links, exposes buried epitopes of proteins, protein unfolding/refolding
  • Specific protocol 1: citrate buffer [1][2]
  • Specific protocol 2: Tris-EDTA buffer [3]

Protease treatment

  • Specific protocol 1: proteinase K [4]
  • Specific protocol 2: trypsin [5]

Detergent treatment

  • Specific protocol: SDS [6]

Personal and lab-specific protocols

  • Korey Griffin's notes:

See also

External links

Commercial antigen retrieval kits: