Characterization of human CD30 overexpressing in 293T stable clones by FACS.
CD30 Cell Line Background
CD30, also known as Ki-1 and TNFRSF8, is a transmembrane protein of TNF receptor superfamily. CD30 is mainly expressed on antigen-stimulated Th cells and B cells in physiological condition, playing a key role in antigen-induced Th cell proliferation and cytokine secretion.
CD30 is also highly expressed in Hodgkin’s disease and other lymphomas, making it a biomarker and drug target. Its monoclonal antibody, Brentuximab, has been approved for tumor treatment.
The TNF receptor superfamily member 8 (CD30) is also known as TNFRSF8, Ki-1, or D1S166E.
Recombinant Cell Line Generation
This 293T human CD30 Cell Line was generated using pLVX-puro vector expressing human CD30 sequence based on an in-house 293T/NF-κb reporter cell line.
Cell Line Characterization
Recombinant Cell Line Profile
|Cell line ID:||293T/hCD30-luc2|
|Cell type:||Human embryonic kidney|
|Gene ID/Accession #:||NM_001243.4|
|Selection marker:||Puromycin (0.5ug/ml) + Hygromycin (50ug/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|
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.