Antigen retrieval

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(Types of retrieval methods)
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== Types of retrieval methods ==
== Types of retrieval methods ==
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# Heat treatment - mechanism: heat cleaves cross-links, exposes buried epitopes of proteins
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=== Heat treatment ===
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# Protease treatment
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* Mechanism: heat cleaves cross-links, exposes buried epitopes of proteins, protein unfolding/refolding
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# Detergent treatment
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* Specific protocol 1: citrate buffer [http://openwetware.org/wiki/Livesey:_Antigen_retrival][http://www.ihcworld.com/_protocols/epitope_retrieval/citrate_buffer.htm]
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* Specific protocol 2: Tris-EDTA buffer [http://www.ihcworld.com/_protocols/epitope_retrieval/tris_edta.htm]
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=== Protease treatment ===
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* Specific protocol 1: proteinase K [http://www.ihcworld.com/_protocols/epitope_retrieval/proteinase-k.htm]
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* Specific protocol 2: trypsin [http://www.ihcworld.com/_protocols/epitope_retrieval/trypsin.htm]
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=== Detergent treatment ===
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* Specific protocol: SDS [http://openwetware.org/wiki/Griffin:Immunofluorescence_Cell_Staining#Antigen_retrieval_for_cryostat_tissue_sections_or_cultured_cells]
== Personal and lab-specific protocols ==
== Personal and lab-specific protocols ==

Revision as of 11:40, 19 October 2009

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.

Contents

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:

Personal tools