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

Name: CEA (Carcinoembryonic Antigen) Antibody

Clone: Rabbit anti-Human

Supplier: Biocare Medical

Catalog Number: CP009B

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.

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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

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


Not need to perform antigen retrieval for acetone fixed frozen sections