Characterization of EGFR mutants overexpressing in BA/F3 stable clones by FACS.
Stable Ba/F3 clone expressing exogenous EGFR isoform a bearing del19 and T790M mutation.
|Cell line ID:||Ba/F3(EGFR)|
|Cell type:||Pro-B cells|
|Gene ID/Accession #:||NM_005228.3|
|Application:||Drug screening and biological assays|
|Morphology:||Mostly single, round (some polymorph) cells in suspension|
|Medium:||RPMI 1640 + 10% FBS|
|Subculture:||Split saturated culture 1:10 every 3 days; seed out at about 1-3 x 105 cells/ml|
|Incubation:||37 °C with 5% CO2|
|Storage:||Frozen in liquid nitrogen with 70% medium, 20% FBS and 10% DMSO|
|Doubling time:||Approximately 20 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.