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Antigen Retrieval for Frozen Sections Protocol

This protocol is intended as a guide only.


1% Sodium Dodecyl Sulfate (SDS) in PBS:

SDS -------------------------------------- 1 g

0.01M PBS (pH 7.4) ---------------- 100 mL

Mix to dissolve.

  1. Rinse sections three times for 5 min each in PBS.
  2. Cover sections with 1% SDS solution and incubate for 5 minutes at RT.
  3. Rinse 3 X 5 min each in PBS. (It is important to wash sections well).
  4. Proceed with staining as normal.


En Bloc Antigen Retrieval:

  1. Fix tissue with buffered 4% PFA. The tissue blocks should be cut to a proper size (e.g. slides 3-5 mm thick).
  2. Immerse the tissue blocks in Citrate Buffer at 4 °C overnight.
  3. Place the tissue blocks at 95-100 °C for 3-5 minutes.
  4. Immediately place the tissue blocks in cold 30% sucrose in PBS and incubate at 4 °C until the blocks sink.
  5. Immerse the tissue blocks in an embedding medium and freeze quickly with crushed dry ice.
  6. The frozen tissue blocks can now be stored at -80 degrees Celsius and process as normal.

* The above information is intended as a guide. Determining the protocol that best meets your need should be determined for each antigen and antibody.