Carcinoembryonic Antigen (CEA) Antibody Staining Protocol for Immunohistochemistry


Description: Carcinoembryonic Antigen (CEA) is an antigen (protein) present in very small quantities in adult tissue. A greater than normal amount may be suggestive of cancer. CEA is therefore called a tumor marker. Tests for its presence aid in screening, in evaluating recurrent or disseminated disease, and in gauging the success of surgical removal of malignant tumors.

Primary Antibody

Name: CEA (Carcinoembryonic Antigen) Antibody

Clone: II-7, Mouse anti-Human

Supplier: Dako

Catalog Number: M7072

Dilution: 1:400 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