FACS analysis of cell surface CD86 expression by an anti-human CD86 antibody and isotype control antibody.
HEK 293T cells stably expressing exogenous human CD86 gene.
|Cell line ID:||293T/CD86|
|Cell type:||Human embryonic kidney|
|Gene ID/Accession #:||NM_175862.4|
|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|
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.