IL-4 & IL-13 Reporter HEK 293 Cells

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IL-4 & IL-13 responsive STAT6-SEAP reporter assay

Signaling pathway in HEK-Blue™ IL-4/IL-13 cells

HEK-Blue™ IL-4/IL-13 cells are designed to monitor human IL-13-induced STAT6 stimulation or inhibition. This colorimetric bioassay can be used for screening activatory molecules, such as engineered cytokines, or inhibitory molecules, such as neutralizing antibodies.

HEK-Blue™ IL-4/IL-13 cells respond specifically to recombinant human IL-4 and recombinant human IL-13. The reliable and consistent performance of HEK-Blue™ IL-4/IL-13 cells makes them suitable for release assays of therapeutic molecules that inhibit IL-4 or IL-13 signaling, such as Dupilumab, a therapeutic monoclonal antibody targeting the common IL-4Rα subunit of the IL-4  and IL-13 receptors (see figures).
 

Key features

• Readily assessable STAT6-SEAP reporter activity
• Convenient readout using QUANTI-Blue™ Solution
• High sensitivity to human (h) IL-4 and hIL-13
• Stability guaranteed for 20 passages

Applications

• Therapeutic development
• Drug screening
• Release assay

IL-4 and IL-13 are cytokines that play an important role in the regulation of inflammation and immune responses.

 More details

InvivoGen’s products are for research use only and not for clinical or veterinary use.

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®

Specificity: Detects human IL-4 and human IL-13

Detection range:

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

Quality Control:

• SEAP reporter activity in response to IL-4 and IL-13 is validated using functional assays.
• The stability for 20 passages following thawing is confirmed.
• These cells are tested for mycoplasma contamination. 

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 and some areas in Asia)

Details

Cell line description

HEK-Blue™ IL-4/IL-13 cells were generated by stable transfection of the human embryonic kidney HEK293 cell line with the gene encoding human STAT6 to obtain a fully active STAT6 pathway. The other genes of the IL-4/IL-13 pathway are naturally expressed in these cells. These cells were further transfected with a STAT6-inducible secreted embryonic alkaline phosphatase (SEAP) reporter. The binding of IL-4 or IL-13 to their receptor triggers a signaling cascade leading to STAT6 activation and the subsequent production of SEAP. This can be readily assessed in the supernatant using QUANTI-Blue™ Solution, a SEAP detection reagent.

HEK-Blue™  IL-4/IL-13 cells detect human (h) IL-4 and hIL-13 (see figures). Of note, they do not respond to hIFN-α or hIFN-β (see figures).

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