Characterization of cynomolgus monkey B7X overexpression in 293T stable clones by FACS.
B7X Cell Line Background
B7X belongs to the B7 costimulatory protein family. Proteins in this family are present on the surface of antigen-presenting cells and interact with ligand bound to receptors on the surface of T cells. Studies have shown that high levels of the encoded protein has been correlated with tumor progression. A pseudogene of this gene is located on chromosome 20. Multiple transcript variants encoding different isoforms have been found for this gene.
Cell Line Generation
293T Cynomolgus monkey B7X cell Line was generated using lentiviral vector expressing monkey B7X plus flag sequence.
Cell Line Characterization
Recombinant Cell Line Profile
|Cell line ID:||293T/Monkey B7X|
|Cell type:||Human embryonic kidney|
|Gene ID/Accession #:||XM_015430437.1|
|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:||Exp. Cell Res. 306:128-141(2005)|
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.