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Carcinoembryonic Antigen (CEA) Antibody Staining Protocol for Immunohistochemistry
Description: Reacts with CEA and CEA-like proteins such as NCA (non-specific cross-reacting antigen) and NCA2 and biliary glycoprotein (BGP1). In all tissues, the NCA of neutrophil granulocytes are stained positive. In colon adenocarcinoma, the antibody labels the cytoplasm of the tumor cells strongly. In the normal colon, the luminal part of enterocytes is strongly labelled. In breast carcinoma, a low percentage of glandular epithelial cells are stained, and gland lumina and some epithelial cell membranes in the normal breast tissue show positive staining. In the liver, the biliary canaliculi are positive. In general, CEA will mark adenocarcinoma of the stomach, colon, lung and pancreas; CEA is weakly or occasionally positive (less than 10%) for prostate cancer, bladder cancer and hepatoma; and CEA is negative for squamous cell carcinoma of the skin and esophagus, mesothelioma, lymphoma, melanoma and sarcoma.
Primary Antibody
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Name: CEA (Carcinoembryonic Antigen) Antibody |
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Clone: Rabbit anti-Human |
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Supplier: Biocare Medical |
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Catalog Number: CP009B |
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Dilution: 1:200 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed. |
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Incubation Time/Temp: 60 min/room temperature |
Antigen Retrieval
| Device: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102) |
| Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100) |
| Heat/Cool Temperature: 95-100 ºC/room temperature |
| Heat/Cool Time: 20 minutes/20 minutes |
Detection Methods
| Standard Method: ABC Method or LSAB Method |
| Enhanced Method: Polymeric Methods |
Chromogen Substrate
| Reagent: DAB |
| Incubation Time/Temperature: 1-3 minutes/room temperature |
Counterstain
| Reagent: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Cytoplasmic |
| Images: Search image |
Additional Information:
| Tissue Type: Colon adenocarcinoma, breast carcinoma |
| Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections |
| Positive Control: Colon adenocarcinoma, breast carcinoma |
| Negative Control: Omit primary antibody, isotype control, absorption control |
| Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary |
Notes:
Not need to perform antigen retrieval for acetone fixed frozen sections |
References: