BaF3 EGFR-H773_V774insNPH BioVector NTCC质粒载体菌种细胞基因保藏中心

BaF3 EGFR-H773_V774insNPH BioVector NTCC质粒载体菌种细胞基因保藏中心 Cell line ID: Ba/F3(EGFR-H773_V774insNPH)
Species: Mouse
Cell type: Pro-B cells
Description: Stable Ba/F3 clone expressing exogenous EGFR isoform a bearing H773_V774insNPH
mutation
Gene ID/Accession #: NM_005228.3
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:
References:
1
Nature 309:418-425(1984)
Background
EGFR (epidermal growth factor receptor) is a cell-surface receptor tyrosine kinase. Activation of this receptor has
been associated with a number of cancers, including lung and colon cancer. The identification of EGFR as a driver
gene has led to the development of anticancer therapeutic agents, including gefitinib, erlotinib, afatinib, osimertinib
(AZD9291), and cetuximab. Ba/F3, a murine interleukin-3 dependent pro-B cell line, is a popular system for exploring
both kinases and their kinase inhibitors, because some protein kinases can render Ba/F3 cells independent of IL-3,
while their inhibitors antagonize this effect.Cell Line Generation
Ba/F3 EGFR-H773_V774INSNPH Cell Line was generated using Retrovirus system expressing EGFR-H773_V774insNPH
sequence.Applications
A. cell-based kinase inhibition screen
B. cell viability assay
C. in vivo efficacy study. 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