IL-4 & IL-13 Reporter HEK 293 Cells

IL-4 & IL-13 Reporter Cells

HEK-Blue™ IL-4/IL-13 Cells signaling pathway

HEK-Blue™ IL-4/IL-13 cells allow the detection of bioactive human interleukin-4 (IL-4) and human/murine IL-13 by monitoring the activation of the STAT-6 pathway. IL-4 and IL-13 are cytokines that play an important role in regulating inflammation and immune responses [1].

 More details

Cell line description:

HEK-Blue™ IL-4/IL-13 cells were generated by stable transfection of the human embryonic kidney HEK 293 cell line with the human STAT6 gene to obtain a fully active STAT6 pathway. The other genes of the pathway are naturally expressed in sufficient amounts. These cells were further transfected with a STAT6-inducible SEAP reporter gene.

HEK-Blue™ IL-4/IL-13 cells produce SEAP in response to IL-4 or IL-13 stimulation. The levels of STAT6-induced SEAP secreted in the supernatant can be easily determined using QUANTI-Blue™ Solution.

Stimulation of HEK-Blue™ IL-4/IL-13 cells with recombinant human IL-4 or IL-13 can be blocked by the neutralizing monoclonal hIL-4 and hIL-13-IgG antibodies, respectively.

Features of HEK-Blue™ IL-4/IL-13 cells:

• Fully functional IL-4/IL-13 signaling pathway
• Readily assessable SEAP reporter activity
• Functionally tested and guaranteed mycoplasma-free

Applications of HEK-Blue™ IL-4/IL-13 cells:

• Detection of human IL-4 and human/murine IL-13
• Screening of anti-IL-4, anti-IL4Rα, anti-IL-13, or anti-IL-13Rα1 antibodies
• Screening of JAK1 inhibitors

Reference:

1. Kasaian MT. et al., 2013. An IL-4/IL-13 dual antagonist reduces lung inflammation, airway hyperresponsiveness, and IgE production in mice. Am J Respir Cell Mol Biol.49(1):37-46.

Figures

Stimulation of HEK-Blue™ IL-4/IL-13 cells by recombinant human IL-4 or IL-13. After a 24 h incubation, SEAP activity was assessed using QUANTI-Blue™ Solution and reading the optical density (O.D.) at 655 nm.

Ligand	EC50	Response Ratio
Recombinant hIL-4	~0.7 ng/ml	25
Recombinant hIL-13	~2.0 ng/ml	28

HEK-Blue™ IL-4/IL-13 cells were stimulated with various human recombinant cytokines; IFN-α (1000 IU/ml), IFN-β (1000 IU/ml), IFN-γ (1000 IU/ml), IL-1β (100 ng/ml), IL-4 (10 ng/ml), IL-6 (100 ng/ml), IL-13 (10 ng/ml), IL-18 (100 ng/ml), TGF-β (10 ng/ml), and TNF-α (100 ng/ml). After a 24 h incubation, SEAP activity was assessed using QUANTI-Blue™ Solution and reading the O.D. at 655 nm.

Specifications

Antibiotic resistance: blasticidin, Zeocin™

Growth medium: DMEM, 4.5 g/l glucose, 2-4 mM L-glutamine, 10% (v/v) heat-inactivated fetal bovine serum, 100 U/ml penicillin, 100 μg/ml streptomycin, 100 μg/ml Normocin™

Guaranteed mycoplasma-free

Detects human IL-4 and human/murine  IL-13

• Detection range for human IL-4: 0.1 - 10 ng/ml
• Detection range for human IL-13: 1 - 100 ng/ml

Contents

• 1 vial containing 3-7 x 10⁶ cells
• 1 ml of Blasticidin (10 mg/ml)
• 1 ml of Zeocin™ (100 mg/ml)
• 1 ml of Normocin™ (50 mg/ml)
• 1 ml of QB reagent and 1 ml of QB buffer (sufficient to prepare 100 ml of QUANTI-Blue™ Solution, a SEAP detection reagent)

Shipped on dry ice (Europe, USA & Canada)

Details

The transcription factor STAT6 is activated primarily by two cytokines with overlapping biologic functions, IL-4 and IL-13 [1, 2].

It can also be activated by IFN-α in a cell-specific manner. In non-hematopoietic cells, IL-4 and IL-13 bind a receptor complex composed of the IL-4Ralpha and IL-13Ralpha1.

Upon ligand binding, the receptor complex activates the receptor-associated Janus kinases (JAK1 and Tyk2) leading to the recruitment of STAT6 and its phosphorylation.

Activated STAT6 forms homodimers that translocate to the nucleus where they bind the promoter of responsive genes inducing gene transcription.

In HEK-Blue™ IL-4/IL-13 cells stimulation with IL-4 or IL-13 activates the JAK/STAT6 pathway with the subsequent production of SEAP.

1. Kasaian MT. et al., 2013. An IL-4/IL-13 dual antagonist reduces lung inflammation, airway hyperresponsiveness, and IgE production in mice. Am J Respir Cell Mol Biol.49(1):37-46.
2. David M. et al., 2003. Functional characterization of IL-13 receptor a2 gene promoter: a critical role of the transcription factor STAT6 for regulated expression. Oncogene 22, 3386–3394.

FAQ

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