Human IL-1α & IL-1β Reporter HEK 293 Cells

Cytokine offer: For each cytokine reporter cell line purchased, get a free vial of the matching cytokine. View all InvivoGen's Cytokine Bioassays.

Human IL-1 Reporter Cells

Signaling pathway in HEK-Blue™ IL-1β cells

HEK-Blue™ IL-1β cells are designed to monitor human IL-1β-induced NF-kB/AP-1 stimulation or inhibition. This colorimetric bioassay can be used for screening activatory molecules, such as engineered cytokines, or inhibitory molecules, such as neutralizing antibodies. These cells can be used together with our Inflammasome test cells to monitor IL-1β secretion in inflammasome activation studies. 

HEK-Blue™ IL-1β cells respond to recombinant human IL-1β and IL-1α. The respond poorly to mouse IL-1β and IL-1α. The reliable and consistent performance of HEK-Blue™ IL-1β cells makes them suitable for release assays of therapeutic molecules that inhibit IL-1 signaling, such as Nidanilimab/Nadunolimab, a monoclonal antibody targeting the human IL-1 receptor accessory protein (IL1RAP or IL1R3) (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-1α and hIL-1β
• Stability guaranteed for 20 passages

Applications

• Detection of IL-1β in inflammasome studies using the THP-1/HEK-Blue™ IL-1β Assay (see below)
• Therapeutic development
• Drug screening
• Release assay

Both IL‑1α and IL-1β are secreted pro‑inflammatory cytokines that play a critical role in immune responses and inflammation [1].

More details

Note: This cell line has been replaced with another clone, which is KO for TLR3, TLR5, and TNFR1. Thus, it has a new cat. code: hkb-il1bv2.

Figures

Stimulation of HEK-Blue™ IL-1β cells by recombinant human and murine IL-1β. After a 24 h incubation, SEAP activity was assessed using QUANTI-Blue™ Solution and reading the optical density (O.D.) at 630 nm.

Stimulation of HEK-Blue™ IL-1β cells by recombinant human and murine IL-1α. After a 24 h incubation, SEAP activity was assessed using QUANTI-Blue™ Solution and reading the optical density (O.D.) at 630 nm.

Specific response of HEK‑Blue™ IL ‑1β cells to IL‑1β and IL‑ 1α cytokines.

Cells were stimulated with 1 ng/ml of hIL‑1β, mIL-1β, hIL-1α, mIL-1α, 100 ng/ml of hTNF-α, mTNF-α, ultrapure flagellin from S. typhimurium (FLA-ST UP) or 300 ng/ml of Poly(I:C) HMW. After overnight incubation, SEAP activity was assessed using QUANTI‑Blue™ Solution. OD at 630 nm is shown as mean ± SEM.

Dose-dependent inhibition of HEK-Blue™ IL-1β cell response using Nidanilimab biosimilar. Increasing concentrations of Anti-hIL1RAP-hIgG1 (0.1 ng/ml - 10 µg/ml) were incubated with HEK‑Blue™ IL-1β cells for 1 hour before the addition of CHO-derived recombinant human IL-1β (100 pg/ml). After overnight incubation, SEAP activity in the cell culture supernatant was assessed using QUANTI-Blue™ Solution. Data are shown as the percentage of activity (mean ± SEM).

Specifications

Cell type: Epithelial

Tissue origin: Human Embryonic Kidney

Target: IL-1α, IL-1β

Specificity: Human

Reporter gene: SEAP

Antibiotic resistance: Zeocin®

Detection ranges:

• Human IL-1β: 100 pg - 100 ng/ml
• Mouse IL-1β: 10 ng - 1 µg/ml

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-1β cells (hkb-il1bv2)

• 1 vial containing 3-5 x 10⁶ cells
• 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-1β vial (hkb-il1bv2-av)

• 1 vial containing 3-5 x 10⁶ cells

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

Notification:  Reference #hkb-il1bv2-av can only be ordered together with reference #hkb-il1bv2.

Details

Cell line description

HEK-Blue™ IL-1β cells were generated by stable transfection of the human embryonic kidney HEK293 cell line. These cells endogenously express the human (h) IL-1 receptor, which binds both IL-1α and IL-1β. They were transfected with an NF-κB/AP-1-inducible secreted embryonic alkaline phosphatase (SEAP) reporter. The binding of IL-1α or IL-1β to its receptor triggers a signaling cascade leading to NF-κB/AP-1 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-1β cells respond to hIL-1α and hIL-1β. They also respond to a lesser extent to mouse (m) IL-1α and mIL-1β. Of note, these cells do not respond to hTNF-α or mTNF-α.

IL-1β background

IL-1β is a soluble pro-inflammatory cytokine that plays a critical role in the host response to infection and injury [1]. It is synthesized as a pro-IL-1β zymogen by activated macrophages and must be cleaved by caspase-1 to generate its mature form [2]. IL-1β binding to the IL-1R1 receptor triggers the formation of the IL-1R1/IL-1R3/MyD88 complex and induces signaling leading to the activation of the transcription factors NK-κB and AP-1 [3]. Due to its role in mediating acute and chronic inflammation, IL-1β has emerged as a therapeutic target for auto-inflammatory diseases [1,4]. 

1. Dinarello C., 2018. Overview of the IL-1 family in innate inflammation and acquired immunity. Immunol Rev. 281(1): 8–27. 
2. Lopez-Castejon G. & Brough D., 2011. Understanding the mechanism of IL-1β secretion. Cytokine Growth Factor Rev. 22(4):189-95.
3. Weber A. et al., 2010. Interleukin-1 (IL-1) pathway. Sci Signal. 3(105):cm1.
4. Dinarello CA., 2011. Interleukin-1 in the pathogenesis and treatment of inflammatory diseases. Blood. 117:3720–3732.

THP-1/HEK-Blue™ IL-1β Assay

THP-1/HEK-Blue™ IL-1β Assay

1. Production of IL-1β by THP-1 cells. Typically, THP-1 cells are pre-treated with phorbol 12-myristate acetate (PMA) to become more susceptible to inflammasome activators, then are primed with lipopolysaccharide (LPS). These treatments induce the production of pro-IL-1β, the immature form of IL-1β. Subsequent stimulation with inflammasome inducers, such as ATP, leads to NRLP3 and caspase-1 activation resulting in IL-1β maturation and secretion.

2. Detection of IL-1β by HEK-Blue™ IL-1β cells. IL-1β-containing THP-1 supernatants are added to HEK-Blue™ IL-1β cells leading to NF-κB activation and the subsequent production of SEAP. The presence of SEAP in HEK-Blue™ IL-1β supernatants is assessed using QUANTI-Blue™ Solution, a SEAP detection medium.

FAQ Cell Lines

Any questions about our cell lines ? Visit our frequently asked questions page