IL-11 Reporter HEK 293 Cells

Interleukin-11 Reporter Cells

Signaling pathway in HEK-Blue™ IL-11 cells

HEK-Blue™ IL-11 cells are designed to monitor human IL-11-induced STAT3 stimulation or inhibition through SEAP detection. This colorimetric bioassay can be used for screening activatory molecules, such as engineered cytokines, or inhibitory molecules, such as neutralizing antibodies.

HEK-Blue™ IL-11 cells respond strongly to recombinant human IL-11 and mouse (m)IL-10 (see figure). The reliable and consistent performance of HEK-Blue™ IL-10 cells makes them suitable for release assays of therapeutic molecules that inhibit IL-11 signaling, such as monoclonal antibodies.

Key Features

• Readily assessable STAT3-inducible SEAP reporter activity
• Convenient readout using  QUANTI-Blue™ Solution
• Strong response to human (h) and mouse (m) IL-11
• Stability guaranteed for 20 passages

Applications

• Therapeutic development
• Drug screening
• Release assay

The IL-11 cytokine belongs to the IL-6 family and has pleiotropic physiological functions, ranging from development and tissue homeostasis to immune responses. 

 More details

Note: A new clone is provided with an improved IL-11 response. The cat code has been changed accordingly (hkb-il11rv2).

Figures

Human IL-11Rα mRNA expression in HEK-Blue™ IL-11 cells.
Total mRNA was extracted from ~1x10⁶ HEK-Blue™ parental cells (blue) and HEK-Blue™ IL-11 cells (red). Human IL-11Rα mRNA was amplified using quantitative RT-qPCR. Data are represented as the log2 fold change comparing hIL-11Rα relative expression between the two cell lines.

Dose-response of HEK-Blue™ IL-11 cells to recombinant IL-11. Cells were stimulated with increasing concentrations of recombinant human IL-11 (hIL-11) and mouse IL-11 (mIL-11). After overnight incubation, the STAT3-induced SEAP activity was determined using QUANTI-Blue™, a SEAP detection reagent. Data are shown as optical density (OD) at 650 nm (mean ± SEM)

Response of HEK-Blue™ IL-11 cells to a panel of cytokines. Cells were stimulated with various human and mouse recombinant cytokines: 10 ng/ml of hIL-11 and mIL-11, hIL-6, hIL-27, hIL-31, and hIFN-γ. After overnight incubation, SEAP activity was assessed using QUANTI-Blue™. Data are shown as optical density (OD) at 650 nm (mean ± SEM).

Specifications

Cell type: Epithelial

Tissue origin: Human Embryonic Kidney

Target: IL-11

Specificity: Human, mouse

Reporter gene: SEAP

Antibiotic resistance: Blasticidin, Hygromycin, Zeocin®

Detection ranges: 

• Human IL-11: 0.1 ng/ml - 100 ng/ml
• Mouse IL-11: 0.1 ng/ml - 100 ng/ml

Growth medium: Complete DMEM (see TDS)

Growth properties: Adherent

Mycoplasma-free: Verified using Plasmotest™

Quality control: Each lot is functionally tested and validated. 

Additional information: The expression of human IL-11Rα is confirmed by RT-qPCR.

Contents

HEK-Blue™ IL-11 Cells (hkb-il11rv2)

• 1 vial containing 3-7 x 10⁶ cells
• 1 ml Normocin® (50 mg/ml)
• 2 x 1 ml of HEK-Blue Selection (250X)
• 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-11 vial (hkb-il11r-av)

• 1 vial containing 3-7 x 10⁶ cells

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

Notification:  Reference #hkb-hil11r-av can only be ordered together with reference #hkb-hil11rv2.

Details

Cell line description

HEK-Blue™ IL-11 cells were generated by stable transfection with the genes encoding the human IL-11 receptor (IL-11Rα and gp130 chains), human STAT3, and a STAT3-inducible secreted embryonic alkaline phosphatase (SEAP) reporter. The binding of IL-11 to its receptor triggers a signaling cascade leading to the activation of STAT3 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-11 cells detect human and murine IL-11. Of note, these cells also respond to human IL-6 and IL-27 as they express the gp130 co-receptor shared by IL-11 and other cytokines of the IL-6 family (see figures). 

IL-11 background

The cytokine interleukin 11 (IL-11) belongs to the IL-6 family,  which also includes IL-6, IL-27 p28/IL-30, IL-31, Leukemia inhibitory factor (LIF), Oncostatin M (OSM), Cardiotrophin-like cytokine (CLC), Ciliary neurotrophic factor (CNTF), Cardiotrophin-1 (CT-1), and Neuropoietin [1-3]. IL-11 has pleiotropic physiological functions, ranging from development and tissue homeostasis, to immune responses [1, 2]. IL-11 signalisation requires a multimeric structure comprising two specific α chains (IL-11Rα), two gp130 chains (common to the IL-6 signaling receptor), and 2 cytokines. This hexameric complex allows the juxtaposition of the intracellular Janus kinases (JAK) and activation of the signal transducer and activator of transcription 3 (STAT3) [1,3]. Other signaling pathways include the Ras-MAPK pathway, the PI3-K-Akt pathway, and the p38 and JNK MAPK pathways [2, 3].

While gp130 is ubiquitously expressed, IL-11Rα expression is cell-specific. Soluble IL-11Rα resulting from IL-11R proteolysis allows trans-signaling and widens the number of target cells [4]. Importantly, IL-11, like IL-6, has no affinity toward gp130 in the absence of its specific receptor α chain [1]. 

1. Garbers C. & Scheller J., 2013. Interleukin-6 and interkeukin-11: Same same but different. Biol. Chem. 394:1145.
2. Kang S. et al., 2020. Historical overview of the interleukin-6 family cytokine. J. Exp. Med. 217: e20190347.
3. Morris R. et al., 2018. The molecular details of cytokine signaling via the JAK/STAT pathway. Protein Science. 27:1984.
4. Lamertz L. et al., 2018. Soluble gp130 prevents interleukin-6 and interleukin-11 cluster signaling but not intracellular autocrine responses. Science Sign. 11(550):eaar7388).

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