14-0669-82

antibody from Invitrogen Antibodies

Targeting: CEACAM5 CD66e, CEA

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

14-0669-82

Provider

Invitrogen Antibodies

Product name

CD66e (CEA) Monoclonal Antibody (CB30), eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: The monoclonal antibody CB30 recognizes human CD66e, also known as CEA (carcinoembryonic antigen) and CEACAM5. As a member of the immunoglobulin superfamily, it is large GPI-linked glycosylated protein with a molecular weight about 200 kDa. It is expressed mainly on epithelial cells and on a variety of tumors of the gastrointestinal, respiratory and genitourinary tracts. CD66e has been shown to function in cell adhesion and migration via homophilic and heterophilic interactions with NCA (non-specific cross-reacting antigen also known as CEACAM6 and CD66c). Overexpression is correlated with tumor invasiveness and is therefore used as a clinical marker.

Antibody clone number

CB30

Concentration

0.5 mg/mL

References

Submitted references

Carcinoembryonic antigen gene family: molecular biology and clinical perspectives.

Thompson JA, Grunert F, Zimmermann W
Journal of clinical laboratory analysis 1991;5(5):344-66

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Carcinoembryonic antigen, a human tumor marker, functions as an intercellular adhesion molecule.

Benchimol S, Fuks A, Jothy S, Beauchemin N, Shirota K, Stanners CP
Cell 1989 Apr 21;57(2):327-34

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-CD66e (CEA) Monoclonal Antibody (CB30), eBioscience™ (Product # 14-0669-82) and a 160 kDa band corresponding to Carcinoembryonic antigen-related cell adhesion molecule 5 was observed across cell lines tested. Membrane enriched extracts (30 µg lysate) of MCF7 (Lane 1), HCT 116 (Lane 2), SW480 (Lane 3), HT-29 (Lane 4), HT-29 treated with IL-6, 100 ng/mL, 48 Hrs. (Lane 5), MDA-MB-231 (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 Dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Carcinoembryonic antigen-related cell adhesion molecule 5 was performed using 70% confluent log phase HT-29 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with CD66e (CEA) Monoclonal Antibody (CB30), eBioscience™ (Product # 14-0669-82) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing cell membrane localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60x magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry of formalin-fixed paraffin embedded human infiltrating ductal carcinoma, using 10 µg/mL of Mouse IgG1 Isotype Control Purified (Product # 14-4714-82) (left) or Anti-Human CD66 (CEA) Purified (right) followed by Anti-Mouse Biotin, and DAB visualization. Nuclei are counterstained with hematoxylin.