14-0669-80

antibody from Invitrogen Antibodies

Targeting: CEACAM5 CD66e, CEA

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Data presented on provider website)

Immunohistochemistry (Data presented on provider website)

Flow cytometry (Recommended by provider)

Antibody Data

Product number

14-0669-80

Provider

Invitrogen Antibodies

Product name

Anti-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. Applications Reported: This CB30 antibody has been reported for use in flow cytometric analysis, immunoblotting (WB), and immunohistochemical staining. Applications Tested: This CB30 antibody has been tested by immunohistochemistry and by western blot on cell lysates prepared from MCF7 cells. This antibody can be used at less than or equal to 1 µg/mL for immunoblotting and less than 10 µg/mL for IHC. It is recommended that this antibody be carefully titrated for optimal performance in the assay of interest. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 µm post-manufacturing filtered.

Reactivity

Human, Mouse

Host

Mouse

Isotype

IgG

Antibody clone number

CB30

Vial size

25 µg

Concentration

0.5 mg/mL

Storage

4° C

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

---

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).