The CD66 (carcinoembryonic antigen, CEA, biliary glycoprotein I, BGP-1, CEACAM) immunoglobulin superfamily of genes encode cell adhesion proteins, which are expressed at higher levels in tumorous tissues than in normal tissues. The human CD66 gene family is a diverse set of glycoproteins of epithelial and hematopoietic lineage that comprises 29 genes, which map to chromosome position 19q13.1-q13.2. CD66A, CD66B, CD66C, CD66D, CD66E and CD66F are the best characterized CD66 antigens, and CD66A-D expression upregulates on the surface of granulocytes upon stimulation. Certain CD66 family members mediate homotypic and heterotypic intercellular adhesion events. CD66E, also known as CEA, is a well known tumor marker and a heavily glycosylated GPI-linked cell surface molecule.
Background References
1. Schrewe H et al. Cloning of the complete gene for carcinoembryonic antigen: analysis of its promoter indicates a region conveying cell type-specific expression. Mol Cell Biol 10:2738-2748 (1990).
2. Hirovuki O et al. Preoperative serum CEA level is predictive for patient outcome in case of non-small cell lung cancer, especially squamous cell carcinoma. Yokohama Med 56(5/6):541-546 (2005).
Sequence Similarity
Belongs to the immunoglobulin superfamily. CEA family.
Tissue Specificity
Expressed in columnar epithelial and goblet cells of the colon (at protein level). Found in adenocarcinomas of endodermally derived digestive system epithelium and fetal colon.
Post-translational Modification
Complex immunoreactive glycoprotein with a MW of 180 kDa comprising 60% carbohydrate.
ICC staining Carcino Embryonic Antigen CEA in SK-Br-3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with Carcino Embryonic Antigen CEA polyclonal antibody at a dilution of 1/200 for at least 1 hour at room temperature, washed with PBS. Alexa Fluor™ 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
ICC staining Carcino Embryonic Antigen CEA in SW620 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with Carcino Embryonic Antigen CEA polyclonal antibody at a dilution of 1/200 for at least 1 hour at room temperature, washed with PBS. Alexa Fluor™ 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
Immunohistochemical analysis of paraffin-embedded human liver cancer tissue using anti-Carcino Embryonic Antigen CEA antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (0807-10) at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue using anti-Carcino Embryonic Antigen CEA antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (0807-10) at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
Flow cytometric analysis of Carcino Embryonic Antigen CEA was done on HepG2 cells. The cells were fixed, permeabilized and stained with Carcino Embryonic Antigen CEA antibody at 1/100 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). After incubation of the primary antibody on room temperature for an hour, the cells was stained with a Alexa Fluor™ 488-conjugated goat anti-rabbit IgG Secondary antibody at 1/500 dilution for 30 minutes.
Western blot analysis of Carcino Embryonic Antigen CEA on different lysates with Rabbit anti-Carcino Embryonic Antigen CEA antibody (0807-10) at 1/1,000 dilution.
Lane 1: MCF7 (Human breast cancer cell) cell lysate
Lysates/proteins at 20 µg/Lane. Exposure time: 20 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: 0807-10, 1/5,000 in 5% NFDM/TBST, overnight at 4 ℃ Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 77 kDa Observed band size: 180-200 kDa
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