IHC sample preparation (frozen vs. paraffin-embedded)
IHC sample fixation (formalin vs. alcohol)
IHC epitope retrieval (HIER vs. PIER)
IHC blocking non-specific binding
IHC primary antibody selection
Secondary Antibodies selection
IHC detection methods
IHC application resources
Multicolor ICC/IF staining
View all immunoassay protocols
Antigen Retrieval for Frozen Sections
1% Sodium Dodecyl Sulfate (SDS) in PBS:
SDS -------------------------------------- 1 g
0.01M PBS (pH 7.4) ---------------- 100 ml
Mix to dissolve.
- Rinse sections three times for 5 min each in PBS.
- Cover sections with 1% SDS solution and incubate for 5 minutes at RT.
- Rinse 3 X 5 min each in PBS. (It is important to wash sections well).
- Proceed with staining as normal.
En Bloc Antigen Retrieval:
- Fix tissue with buffered 4% PFA. The tissue blocks should be cut to a proper size (e.g. slides 3-5 mm thick).
- Immerse the tissue blocks in Citrate Buffer at 4 degrees Celsius overnight.
- Place the tissue blocks at 95-100 degrees Celsius for 3-5 minutes.
- Immediately place the tissue blocks in cold 30% sucrose in PBS and incubate at 4 degrees Celsius until the blocks sink.
- Immerse the tissue blocks in an embedding medium and freeze quickly with crushed dry ice.
- 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.