KC-0873

293T-IL4R-Cell-Line

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Background of 293T-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-0873
Cell Line Name	293T-IL4R-Cell-Line
Host Cell Line	Human HEK293T cell line
Description	Stable HEK293T cell line expressing exogenous human IL4Ra & IL13R1 gene
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% DMEM+20% FBS+10% DMSO
Propagation Medium	DMEM+10%FBS+1µg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:3-1:6 every 3 days; seed out at about 1-3 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 30 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

293T human type II IL4R cell line was generated using a plasmid vector expressing human IL4Ra and IL13R1sequence.

Characterization

Figure: Characterization of IL4R overexpressing in the 293T stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (DMEM + 10% FBS + 1μg/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 1:3-1:6 every 3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% DMEM + 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, M., Giallourakis, C. C., Pan, P.-Y., & Rothman, P. B. (2001). Immunoreceptor tyrosine-based inhibitory motif of the IL-4 receptor associates with SH2-containing phosphatases and regulates IL-4-induced proliferation. The Journal of Immunology, 167(11), 6382-6387. https://doi.org/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.