BaF3(SLC34A2-ROS1 BioVector NTCC质粒载体菌种细胞基因保藏中心

BaF3(SLC34A2-ROS1 BioVector NTCC质粒载体菌种细胞基因保藏中心 SLC34A2-ROS1 Cell Line
Catalog # C2066
Cell line ID: Ba/F3(SLC34A2-ROS1)
Species: Mouse
Cell type: Pro-B cells
Description: Stable Ba/F3 clone expressing exogenous SLC34A2-ROS1 fusion protein.
Gene ID/Accession #: EU236946.1
Selection marker: Puromycin
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
Biosafety level: 1
References: Cell 131 (6), 1190-1203 (2007)
Background
ROS1 is a receptor tyrosine kinase (RTK) belonging to the insulin receptor family. Recently, a ROS1 gene rearrangement was
identified as a driver mutation for various cancers, such as glioblstoma tumor, non-small cell lung cancer and
cholangiocarcinoma. The ROS1 protein is structurally similar to the anaplastic lyphoma kinase (ALK) protein, and
the inhibitor crizotinib was also approved for the treatment of NSCLC patients with ROS1 mutation.
Cell Line Generation
Ba/F3 SLC34A2-ROS1 cell line was generated using retrovirus system expressing SLC34A2-ROS1 sequence.Characterization of ROS1 and its mutants overexpressed in Ba/F3 stable clones using Western Blot.
Applications
A. Cell-based kinase inhibition screen
B. Cell viability assay
C. In vivo efficacy study
Example: Kinase inhibitors screening. 1. Harvest and seed the Ba/F3 clones expressing SLC34A2/ROS1 in 96-well plate (3000 cells/90ul medium).
2. Next day, add 10ul 10X serially diluted compound solution each well and incubate the plates for another 72 hours.
3. Add 100ul Cell Titer-Glo each well, mixed and readout using Envision.
4. Plot the dose-responsive curve and fit the IC50 (the concentration of 50% inhibition of DMSO vehicle treated clones)
using GraphPad Prism software (Version 5)
Resuscitation Protocol
Thaw the vial as soon as possible upon receipt to insure its viability.
1. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
2. 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.
3. Spin at ~ 125 x g, for 5- 7 minutes at room temperature.
4. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into
a T25 culture flask.
5. Incubate the culture at 37°, 5% CO2 incubator or refer to the specific culture condition, if possible. BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心www.biovector.net [Supplier来源] http://www.biovector.net