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You are here:Home » Molecular Biology » MB-Cancer Markers » CEACAM 6, Recombinant, Human (Carcinoembryonic Antigen Related Cell Adhesion Molecule)

CEACAM 6, Recombinant, Human (Carcinoembryonic Antigen Related Cell Adhesion Molecule)

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Specifications

Carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM-6), previously called nonspecific crossreacting antigen (NCA) or CD66c, is one of seven human CEACAM family members within the immunoglobulin superfamily (1-4). In humans, CEACAMs include type I transmembrane proteins (CEACAM-1, -3, and -4) and GPI-linked molecules (CEACAM-5 through -8) (1). There is no human CEACAM-2. Human CEACAM-6 is a 90 kD, GPI-linked membrane protein that contains a 34 amino acid (aa) signal sequence, a 286 aa extracellular domain (ECD), and a 24 aa hydrophobic C-terminal propeptide. The GPI membrane anchor is attached at the C terminus following cleavage of the propeptide. CEACAM-6 contains one N-terminal V-type Ig-like domain (N domain), followed by two C2-type Ig-like domains (2-4). It shows considerable glycosylation, including (sialyl) Lewis X , which mediates binding to E-selectin, galectins and some bacterial fimbrae (1, 2). Mature human CEACAM-6 shows 84%, 85%, 80%, 87% and 51% aa identity to the equivalent extracellular regions of human CEACAMs 1, 5 (CEA) and 8, rhesus CEACAM-2, and bovine CEACAM-6, respectively. CEACAM-6 is expressed by granulocytes and their precursors. Activation enhances surface expression by mobilizing CEACAM-6 from storage in azurophilic granules (5, 6). It often shows aberrant expression in acute lymphocytic leukemias (10). CEACAM-6 is also expressed epithelia of various organs and is upregulated in pancreatic and colon adenocarcinomas and hyperplastic polyps (7, 8). Over-expression confers resistance to adhesion-related apoptosis (anoikis) in tumor cells (8, 9). CEACAM-6 is an intercellular adhesion molecule, forming both homotypic, and heterotypic bonds with CEACAM-1, -5 and -8 through interaction of the V-type Ig-like domains (11, 12). Cross-linking of neutrophil CEACAM-6 augments v 3 and 2 integrin-mediated adhesion, apparently by src and caveolin-mediated inside-out integrin activation (8, 13, 14).
Catalog #C2589-79C
SourceHuman CD33 Signal Peptide (Met 1-Ala 16) Human CEACAM-6 (Lys 35-Gly 320) HHHHHH. A DNA sequence encoding the mature human CEACAM-6 (Lys 35-Gly 320; Accession # P40199) (Barnett, T. et al., 1998, Genomics 3(1):59) was fused to the signal peptide from human CD33 at the amino terminus and to a polyhistidine tag at the carboxy-terminus. The chimeric protein was expressed in a mouse myeloma cell line, NS0.
Molecular MassBased on N-terminal sequencing, the recombinant human CEACAM-6 starts at Lys 35 and has a calculated molecular mass of approximately 32.0kD. As a result of glycosylation, the recombinant protein migrates as an approximately 57-75kD protein in SDS-PAGE under reducing conditions.
Endotoxin Level 1 EU/ug of the protein as determined by the LAL method.
ActivityMeasured by the ability of the immobilized protein to support the adhesion of 1 M calcium ionosphere treated human neutrophils. When 2x10e5 cells/well are added to CEACAM-6-coated plates (10ug/ml, 100ul/well), 35-60% of the cells will adhere after 20 minutes at 37°C.
ReconstitutionIt is recommended that sterile PBS be added to the vial to prepare a working stock solution of no less than 100ug/ml. The carrier-free protein should be used immediately upon reconstitution to avoid losses in activity due to non-specific binding to the inside surface of the vial. For long term storage as a dilute solution, a carrier protein (e.g. 0.1% HSA or BSA) should be added to the vial.
StorageLyophilized samples are stable for up to twelve months at -20°C. Upon reconstitution, this protein, in the presence of a carrier protein, can be stored under sterile conditions at 2-8°C for one month or at -20°C in a manual defrost freezer for three months without detectable loss of activity. Avoid repeated freeze-thaw cycles.
SourceMouse myeloma cell line, NS0
Purity95%, as determined by SDS-PAGE and visualized by silver stain.
ConcentrationAs reported
FormSupplied as a lyophilized powder from a 0.2um filtered solution in PBS.
Important NoteThis product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.


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