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

All InvivoGen products are for internal research use only and not for human or veterinary use.

Figures

Dose-response of HEK-Blue™ IL-4/13 cells to recombinant IL-4 and IL-13. Cells were stimulated with increasing concentrations of recombinant human (h)IL-4 and recombinant hIL-13. After overnight incubation, the STAT6-induced SEAP activity was determined using QUANTI-Blue™, a SEAP detection reagent. Data are shown as optical density (OD) at 650 nm (mean ± SEM).

Dose-dependent inhibition of HEK-Blue™ IL-4/13 cell response using Dupilumab biosimilar. HEK-Blue™ IL-4/IL13 cells were incubated with increasing concentrations of Anti-hIL-4R-hIgG4 (S228P) (0.1 ng/ml - 10 µg/ml) for 1 h before the addition of recombinant human (h)IL-4 (1 ng/ml) or hIL-13 (1 ng/ml). After overnight incubation, SEAP activity in the cell culture supernatant was assessed using QUANTI-Blue™ Solution. Data are shown in percentage of activity (%) (mean ± SEM)

Response of HEK-Blue™ IL-4/13 cells to a panel of cytokines. Cells were stimulated with various human (h) recombinant cytokines: 100 ng/ml of hTNF-α, hIL-1β, hIL-2, hIFN-γ; 1 ng/ml of hIL-4, hIL-13; and 1000 U/ml of hIFN-α and hIFN-β. After overnight incubation, SEAP activity was assessed using QUANTI‑Blue™. The OD at 650 nm is shown as mean ± SEM.

Specifications

Cell type: Epithelial

Tissue origin: Human Embryonic Kidney

Target: IL-4 and IL-13

Specificity: Human

Reporter gene: SEAP

Antibiotic resistance: Blasticidin, Zeocin®

Detection range: 20 pg/ml -100 ng/ml (hIL-4), 0.2 ng/ml - 100 ng/ml (hIL-13)

Growth medium: Complete DMEM (see TDS)

Growth properties: Adherent

Mycoplasma-free: Verified using Plasmotest™

Quality control: Each lot is functionally tested and validated.

Contents

HEK-Blue™ IL-4/IL-13 (hkb-il413-4/-13)

• 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)
   

HEK-Blue™ IL-4/IL-13 vial (hkb-il413-av)

• 1 vial containing 3-7 x 10⁶ cells

Shipped on dry ice (Europe, USA, Canada and some areas in Asia)

Notification:  Reference #hkb-il413-av can only be ordered together with references #hkb-il413-4 and/or #hkb-il413-13.

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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