Characterization of CEACAM-1 variants overexpressed in 293T stable clones using Western Blot.
CEACAM-1 Cell Line Background
CEACAM-1 is a member of the carcinoembryonic antigen (CEA) gene family, which belongs to the immunoglobulin superfamily. CEACAM-1 was originally described in bile ducts of liver as biliary glycoprotein. Subsequently, it was found to be a cell-cell adhesion molecule detected on leukocytes, epithelia, and endothelia. The encoded protein mediates cell adhesion via homophilic as well as heterophilic binding to other proteins of the subgroup. Multiple cellular activities have been attributed to the encoded protein, including roles in the differentiation and arrangement of tissue three-dimensional structure, angiogenesis, apoptosis, tumor suppression, metastasis, and the modulation of innate and adaptive immune responses. Multiple transcript variants encoding different isoforms have been reported, but the full-length nature of all variants has not been defined.
Cell Line Generation
293T Human CEMCAM-1 short isoform cell line was generated using lentiviral vector with human CEACAM-1 gene sequence.
Cell Line Characterization
Recombinant Cell Line Profile
|Cell line ID:||293T/CEACAM-1 short isoform|
|Cell type:||Human embryonic kidney|
|Gene ID/Accession #:||NM_001184816|
|Selection marker:||Puromycin (0.5ug/ml)|
|Application:||Drug screening and biological assays|
|Morphology:||Fibroblastoid cells growing adherently as monolayer|
|Medium:||DMEM + 10% FBS|
|Subculture:||Seed out at ca. 2-3 x 106 cells/80 cm2; split confluent culture 1:4 - 1:5 every 2-3 days|
|Incubation:||37 °C with 5% CO2|
|Storage:||Frozen in liquid nitrogen with 70% medium, 20% FBS and 10% DMSO|
|Doubling time:||24-30 hours|
|References:||Nature Reviews. Immunology. 6 (6): 433–46.|
Thaw the vial as soon as possible upon receipt to insure its viability.
- Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
- Transfer the vial into biosafety cabinet. Wipe the surface with 70% ethanol and drop the cell suspension gently into a centrifuge tube containing 9.0 mL complete culture medium.
- Spin at ~ 125 x g, for 5- 7 minutes at room temperature.
- Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
- Incubate the culture at 37°, 5% CO2 incubator or refer to the specific culture condition, if possible.