Product Specifications for Recombinant Human CEACAM-1/CD66a Protein, CF
Mouse myeloma cell line, NS0-derived human CEACAM-1/CD66a protein Gln35-Gly428, with a C-terminal 10-His tag
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
<0.10 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis
Predicted Molecular Mass
100-130 kDa, reducing conditions
Measured by its ability to inhibit IL-2 secretion by HuT 78 human cutaneous T cell lymphoma cells Chen, C.J. and J.E. Shively (2004) J. Immunol. 172:3544. rhCEACAM-1 at concentrations higher than 1 µg/mL reduces anti-CD3 (immobilized at 10 µg/mL) induced IL-2 production by 60-70%. The ED50 for rhCEACAM-1 inhibition of anti-CD3 induced IL-2 production is 0.05-0.2 µg/mL.
Formulation, Preparation and Storage
What does CF mean?
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our
Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant
protein to be stored at a more dilute concentration.
The carrier free version does not contain BSA.
What formulation is right for me?
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or
as an ELISA standard.
In contrast, the carrier free protein is recommended for applications, in which the presence of BSA
Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitute at 100 μg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
3 months, -20 to -70 °C under sterile conditions after reconstitution.
Carcinoembryonic antigen (CEA)-related cell adhesion molecule 1 (CEACAM-1; also BGP) is a 160 kDa member of the CEACAM branch of the CEA gene family of the immunoglobulin superfamily (1 - 3). It is one of seven human CEACAM subfamily genes that are essentially divided equally between type I transmembrane proteins (CEACAM-1, 3 & 4) and GPI-linked molecules (CEACAM-5-8). There is no CEACAM-2 in human. The gene for human CEACAM-1 codes for a 526 amino acid (aa) type I transmembrane protein that contains a 34 aa signal sequence, a 394 aa extracellular domain (ECD), a 24 aa transmembrane segment, and a 74 aa cytoplasmic region (4, 5). The ECD contains one N-terminal V-type Ig-like domain, followed by three C2-type Ig-like domains. It shows considerable glycosylation, including high mannose residues and (sialyl) LewisX (1). The cytoplasmic region shows one ITIM motif and a calmodulin binding site (1 - 3). In addition to the full length form, ten alternate splice forms have been reported (1, 4, 6, 7, 8). There are three soluble and seven transmembrane isoforms, with variations occurring in both the ECD and cytoplasmic region. All ten alternate splice forms contain the V-type Ig-like domain (aa’s 35 - 142). The three soluble forms also contain the first two C2-type Ig-like domains (aa’s 145 - 317), with differences coming in the third C2-type Ig-like domain (6). The seven transmembrane isoforms are highly divergent. Five of the seven contain the V-type plus the first two C2-type domains and then diverge considerably both in the ECD and cytoplasmic region. The remaining two contain only the V-type Ig-like domain, the transmembrane region, and either a full-length or truncated cytoplasmic tail (1, 8). The actual functions of the isoforms are unclear. Full-length mouse and rat CEACAM-1 are approximately 57% aa identical to human CEACAM-1; in the V-type Ig-like domain, they are 58% and 56% aa identical, respectively. The full-length molecule is found on neutrophils, bile duct epithelium, activated NK cells, colonic columnar epithelium and endothelium. It is known to act as an intercellular adhesion molecule, forming both homotypic, and heterotypic bonds with CEA and CEACAM-6/NCA (3, 9). On neutrophils, CEACAM-1 also binds to dendritic cell CD-SIGN via its LeX moiety, inducing dendritic cell maturation and a subsequent Th1-type response (10,11).
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Thompson, J. et al. (1992) Genomics 12:761.
Waggener, C. and S. Ergun (2000) Exp. Cell Res. 261:19.
Barnett, T.R. et al. (1989) J. Cell Biol. 108:267.
Hinoda, Y. et al. (1988) Proc. Natl. Acad. Sci. USA 85:6959.
Kuroki, M. et al. (1991) Biochem. Biophys. Res. Commun. 176:578.
Barnett, T.R. et al. (1993) Mol. Cell. Biol. 13:1273.
Watt, S.M. et al. (1994) Blood 84:200.
Oikawa, S. et al. (1992) Biochem. Biophys. Res. Commun. 186:881.
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