IL-36 Reporter HEK 293 Cells

NF-κB-SEAP reporter cells

ABOUT

IL-36 responsive NF-κB/AP1-SEAP reporter assay

HEK-Blue™ IL-36 cells are designed to monitor human IL-36-induced NF-κB/AP1 stimulation or inhibition. This colorimetric cytokine bioassay can be used to screen activatory or inhibitory molecules, such as engineered cytokines and neutralizing antibodies.

HEK-Blue™ IL-36 cells respond to recombinant human IL-36α, IL-36β, and  IL-36γ, but not to mouse IL-36 (see figures). Their reliable and consistent performance makes them suitable for release assays of activatory and inhibitory molecules such as Imsidolimab, a monoclonal antibody that targets the IL-36 receptor (IL-36R) and prevents IL-36 signaling (see figures).

 

Key features

  • Readily assessable NF-κB/AP-1-inducible SEAP reporter activity
  • Convenient readout using QUANTI-Blue™ Solution
  • Strong response to human (h) IL-36α and IL-36γ
  • No response to mouse (m) IL-36α
  • Stability guaranteed for 20 passages 

Applications

  • Therapeutic development
  • Drug screening
  • Release assay

 

The interleukin-36 (IL-36) belongs to the IL-1 superfamily and comprises multiple isoforms, whereas IL-36α, IL-36β, and IL-36γ mediate pro-inflammatory functions, while a fourth one, IL-36Ra, acts as an antagonist. Dysregulation of IL-36 isoform expression and signaling has been associated with inflammatory diseases such as psoriasis, rheumatoid arthritis, and inflammatory bowel disease.

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Disclaimer:  These cells are for internal research use only and are covered by a Limited Use License (See Terms and Conditions). Additional rights may be available.

Signaling pathway in HEK-Blue™ IL-36 cells
Signaling pathway in HEK-Blue™ IL-36 cells

SPECIFICATIONS

Specifications

Target

IL-36

Target species

Human

Tested applications

Detection and quantification of IL-36 activity

Cell type
Epithelial
Growth properties
Adherent
Tissue origin
Human embryonic kidney cells
Reporter gene
SEAP
Detection method
Colorimetric
Detection range

30 pg/ml - 100 ng/ml (hIL-36α, hIL-36γ)
10 ng/ml - 100 ng/ml (hIL-36β)

Growth medium

Complete DMEM (see TDS)

Antibiotic resistance
Blasticidin
Zeocin®
Mycoplasma-free

Verified using Plasmotest™

Quality control

Each lot is functionally tested and validated.

CONTENTS

Contents

  • Product: 
    HEK-Blue™ IL-36 Cells
  • Cat code: 
    hkb-hil36r
  • Quantity: 
    3-7 x 10^6 cells
Includes:
  • 1 ml Normocin® (50 mg/ml)
  • 1 ml Blasticidin™ (10 mg/ml)
  • 1 ml Zeocin® (100 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)

Shipping & Storage

  • Shipping method:  Dry ice

    • Storage:

    • Liquid nitrogen vapor
    Stability: 20 passages

    Caution:

    • Upon receipt, store immediately in liquid nitrogen vapor. Do not store cell vials at -80°C.

Details

Cell line description

HEK-Blue™ IL-36 cells were generated by stable transfection with the genes encoding for the human IL-36 heterodimeric receptor, comprising the IL-1R6 (aka IL-36R) and IL-1R3 (aka IL-1RAcP) subunits, and an NF-κB/AP-1-inducible secreted embryonic alkaline phosphatase (SEAP) reporter. The binding of IL-36 to its receptor triggers a signaling cascade leading to the NF-κB/AP-1 activation and the subsequent production of SEAP.
HEK-Blue™ IL-36 cells respond to human (h) but not murine (m) IL-36 agonist isoforms. Of note, they are more sensitive to hIL-36α and hIL-36γ compared to hIL-36β (see figures). HEK-Blue™ IL-36 cells maintain their responses to other cytokines that signal through NF-κB/AP-1, such as hIL-1β and hTNF-α (see figures). 

 

IL-36 background

The cytokine interleukin 36 (IL-36) belongs to the IL-1 superfamily. Three isoforms, IL-36α, IL-36β, and IL-36γ, mediate pro-inflammatory functions, while a fourth one, IL-36Ra, acts as an antagonist [1,2].  IL-36 signalization requires the formation of a complex comprised of two subunits, IL-1R6 (aka IL-36R) and IL-1R3 (aka IL-1RAcP, IL-1 receptor accessory protein). The binding of agonist ligands to the IL-36R allows the recruitment of IL-1RAcP and the production of pro-inflammatory cytokines and chemokines through the activation of NF-κB and AP-1 [1,2]. The IL-36Ra antagonist inhibits the signaling by binding to IL-36R and preventing the recruitment of IL-1RAcP [1,2]. IL-36-associated immune response mainly takes place in barrier tissues, such as the skin, lungs, and intestines. Dysregulation of IL-36 isoform expression and signaling has been associated with inflammatory diseases such as psoriasis, rheumatoid arthritis, and inflammatory bowel disease [1,2].

 

​References:

1. Buhl A-L. & Wenzel J.,  2019. Interleukin-36 in infectious and inflammatory skin diseases. Front. Immunol. 10(1162). doi: 10.3389/fimmu.2019.01162.
2. Zhou L. & Todorovic V.,  2021. Interleukin-36: Structure, Signaling and Function. Protein Reviews: Volume 21. doi: 10.1007/5584_2020_488.

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