KC-6473

CHOK1-rat-FGFR2b-Low-Cell-Line

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Background of CHOK1-rat-FGFR2b-Low-Cell-Line

rat-FGFR2b promotes fibroblast growth factor binding activity. It is involved in various processes, including the fibroblast growth factor receptor signaling pathway, forebrain neuron generation, and RNA polymerase II-mediated positive regulation of transcription. It is located in or acts upstream of multiple processes, including lung development, epithelial morphogenesis, and positive regulation of cell population proliferation. It is localized in excitatory synapses and the cell nucleus. It is expressed in various structures, including the digestive system, brain, urogenital system, sensory organs, and skeleton. It is used to study Beare-Stevenson craniosynostosis syndrome, atopic dermatitis, intestinal atresia, and multiple joint fusions. The human homolog of this gene is associated with various diseases.

Specifications

Catalog Number	KC-6473
Cell Line Name	CHOK1-rat-FGFR2b-Low-Cell-Line
NCBI/UniProt Accession Number	NM_012712.1
Clone Number	9#
Host Cell Line	CHOK1
Description	Stable CHOK1 clone expressing exogenous rat-FGFR2b gene in low level
Quantity	Two vials of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	Drug screening and biological assays
Freezing Medium	70% RPMI 1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI 1640 + 10% FBS + 10μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:6-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Doubling Time	Approximately 24 hours
Mycoplasma Status	Negative

Cell Line Generation

CHOK1-rat-FGFR2b-Low-cell-line was generated using a lentiviral vector expressing the rat-FGFR2b sequence.

Characterization

Figure 1: Characterization of rat-FGFR2b overexpression in the CHOK1-rat-FGFR2b-Low stable clone using FACS.

Figure 2: Characterization of rat-FGFR2b in the CHOK1-rat-FGFR2b-Low stable clone using PCR sequencing.

Figure 3: Characterization of rat-FGFR2b overexpression in the CHOK1 stable clone using FACS.

Cell Resuscitation

1. Prewarm culture medium (RPMI 1640 supplemented with 10% FBS and 10μg/mL puromycin)in a 37°C water bath.
2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
7. Incubate the flask at 37°C, 5% CO₂ incubator.
8. Split saturated culture 1:6-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% RPMI 1640 + 20% FBS + 10% DMSO) fresh immediately before use.
2. Keep the freezing medium on ice and label cryovials.
3. Transfer cells to a sterile, conical centrifuge tube, and count the cells.
4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
5. Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
6. Aliquot 1 mL of the cell suspension into each cryovial.
7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
8. Transfer vials to liquid nitrogen for long-term storage

References

1.Hunter DJ, Kraft P, Jacobs KB, Cox DG, Yeager M, Hankinson SE, Wacholder S, Wang Z, Welch R, Hutchinson A, Wang J, Yu K, Chatterjee N, Orr N, Willett WC, Colditz GA, Ziegler RG, Berg CD, Buys SS, McCarty CA, Feigelson HS, Calle EE, Thun MJ, Hayes RB, Tucker M, Gerhard DS, Fraumeni JF, Hoover RN, Thomas G, Chanock SJ (Jul 2007). "A genome-wide association study identifies alleles in FGFR2 associated with risk of sporadic postmenopausal breast cancer". Nature Genetics.