HEK-Blue™ IL-4/IL-13 Cells

IL-4 and IL-13 Sensor Cells

HEK-Blue™ IL-4/IL-13 cells allow the detection of bioactive human IL-4 and human/mouse IL-13 by monitoring the activation of the STAT-6 pathway. These cells were generated by stable transfection of HEK293 cells with the human STAT6 gene to obtain a fully active STAT6 pathway. The other genes of the pathway are naturally expressed in sufficient amounts. The 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™. 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-IgA and hIL-13-IgG antibodies, respectively.

APPLICATIONS: HEK-Blue™ IL-4/IL-13 cells can be used to validate the functionality of recombinant native or engineered human IL-4 or human IL-13. They can also be used to screen for molecules that inhibit IL-4/IL-13 signaling such as antibodies targeting IL-4, IL-13, IL4Rα or IL-13Rα1 and inhibitors of JAK1.

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™ 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™ 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) fetal bovine serum, 50 U/ml penicillin, 50 μg/ml streptomycin, 100 μg/ml Normocin™

Guaranteed mycoplasma-free

Shipped on dry ice

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

• 100 μl Blasticidin (10 mg/ml)

• 100 μl Zeocin™ (100 mg/ml)

• 1 ml Normocin™ (50 mg/ml)

• 1 pouch of QUANTI-Blue™ (SEAP detection medium)

Description

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