KC-1382

CHOK1-IL4R-Cell-Line

22029

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Background of CHOK1-IL4R-Cell-Line

IL4 receptor complex is composed of IL-4Ra subunit and one accessory protein, the common cytokine receptor chain or IL-13Ra1, both of which transduce signals through IL4Ra. Type I IL-4R complexes, composed of IL-4Ra and common cytokine receptor, dominate in hematopoietic lineages and exclusively bind to IL-4 but not to IL-13. Type II IL-4R complexes are composed of IL-4Ra and IL-13Ra1. They are present on both hematopoietic and nonhematopoietic tissues and bind to both IL-4 and IL-13.

Specifications

Catalog Number	KC-1382
Cell Line Name	CHOK1-IL4R-Cell-Line
NCBI/UniProt Accession Number	NM_000418.4
Host Cell Line	Chinese hamster ovary CHOK1 cell line
Description	CHOK1 cell line stable expressing exogenous human IL4R and IL13Ra genes.
Quantity	One vial 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% RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10%FBS+10µg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-2 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 24 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

CHOK1-IL4R cell line was generated using plasmid vector expressing human IL4R and IL13Ra sequence.

Characterization

Figure: Characterization of IL4R overexpressing in CHOK1 stable clones using FACS.

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 10% FBS + 10µg/mL Puromycin) in a 37°C water bath.
Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
Incubate the flask at 37°C, 5% CO₂ incubator.
Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-2 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI1640 + 20% FBS + 10% DMSO) fresh immediately before use.
Keep the freezing medium on ice and label cryovials.
Transfer cells to a sterile, conical centrifuge tube, and count the cells.
Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
Aliquot 1 mL of the cell suspension into each cryovial.
Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
Transfer vials to liquid nitrogen for long-term storage

References

Kashiwada, Masaki, Cosmas C Giallourakis, Ping-Ying Pan, and Paul B Rothman. 2001. ¡ùImmunoreceptor Tyrosine-Based Inhibitory Motif of the IL-4 Receptor Associates with SH2-Containing Phosphatases and Regulates IL-4-Induced Proliferation.¡ì The Journal of Immunology 167 (11). American Association of Immunologists: 6382Ã¿87. doi:10.4049/jimmunol.167.11.6382.
Ikizawa, Koichi, and Yukiyoshi Yanagihara. 2000. ¡ùPossible Involvement of Shc in IL-4-Induced Germline ÈÖ Transcription in a Human B Cell Line.¡ì Biochemical and Biophysical Research Communications 268 (1): 54Ã¿59. doi:10.1006/bbrc.2000.2080.
Maes, T, G F Joos, GG Brusselle American journal of respiratory cell, 2012. n.d. ¡ùTargeting Interleukin-4 in Asthma: Lost in Translation.¡ì Atsjournals.org

Use License Agreement

Research Use Only.

Not for use in diagnostic procedures or therapeutic applications.

Redistribution of the cell line or its derivatives is prohibited without prior written permission from Kyinno Biotechnology.