Recombinant human IL-6 protein - Bioactive cytokine

Human IL-6 protein - E. coli -expressed, tag-free, with HSA

Recombinant human IL-6 is a high-quality and biologically active cytokine, validated using proprietary IL-6 reporter cells. This gp130 cytokine receptor family member is produced in E. coli and thoroughly purified to remove endotoxins.

Recombinant human IL-6 can be used together with HEK-Blue™ IL-6 cells for the screening of inhibitory molecules, such as Tocilizumab, a therapeutic monoclonal antibody targeting the IL-6Rα subunit of the IL-6 receptor (see figures).

IL-6 signaling and biological functions

Key features

• Each lot is validated using HEK-Blue™ IL-6 cells
• Endotoxin < 0.1 EU/µg
• 0.2 µm sterile-filtered

Applications

• Standard for IL-6 detection and quantification assays
• Screening and release assays for antibodies blocking IL-6 signaling
• Screening and release assays for engineered IL-6

Interleukin 6 (IL-6) is a multifunctional cytokine involved in the generation of acute phase proteins, immune responses against pathogens, inflammation, hematopoiesis, cell differentiation, and metabolism. IL-6 overexpression contributes to several inflammatory pathologies, including autoimmune diseases, cancer development, and obesity.

 More details

InvivoGen’s products are for research use only, and not for clinical or veterinary use.

Figures

SDS PAGE of the recombinant human (h)IL-6 protein. 1.5 μg of hIL-6 was loaded on a 12% Mini-PROTEAN® TGX Stain-Free™ Precast Gel (Bio-Rad) stained with Coomassie. Detection was performed as per the manufacturer’s instructions. A band was detected at ~21 kDa

Dose-response in HEK-Blue™ IL-6 cells to recombinant IL-6 cytokine. Cells were stimulated with increasing concentrations of recombinant human IL-6. After overnight incubation, the NF-κB-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-6 cells response using tocilizumab biosimilar. Anti-hIL6R-To-hIgG1 (0 - 1 μg/ml) was incubated with HEK-Blue™ IL-6 cells for 3 hours. Following this, recombinant human IL-6 (0.3 ng/ml) was added to the cells. After overnight incubation, STAT3‑dependent SEAP activity in the cell culture supernatant was assessed using QUANTI-Blue™ Solution, a SEAP detection reagent. Data are presented as percentage (%) of the maximum response.

Specifications

Source: E. coli

Species: Human

Alternative names: B-cell stimulatory factor 2, CTL differentiation factor, Hybridoma growth factor, Interferon beta-2

Carrier: HSA (human serum albumin)

Tag: Tag-free

Accession number: P05231

Protein size: 183 a.a. (V30-M212)

Molecular weight: ~ 21 kDa (SDS-PAGE)

Solubility: 100 μg/ml in water

Formulation: Phosphate buffer saline (pH 7.4), 5% saccharose, 2% HSA

Sterility: 0.2 µm filtration

Form: Lyophilized

Reconstitution buffer: Endotoxin-free water (provided)

Purity: ≥95% (SDS-PAGE)

Endotoxin level: ≤ 0.01 EU/μg (measurement by kinetic chromogenic LAL assay)

Tested applications: Cellular assays

Quality control: Each lot is functionally tested and validated

Contents

Recombinant human IL-6 is provided as a lyophilized powder and is available in two quantities:

• rcyec-hil6: 10 μg
• rcyec-hil6-5: 50 μg (5 x 10 μg)
• 1.5 ml endotoxin-free water for rcyec-hil6 and rcyec-hil6-5

 Recombinant human IL-6 is shipped at room temperature.

Upon receipt, the product should be stored at -20°C.

 Avoid repeated freeze-thaw cycles.

Details

The proinflammatory cytokine IL-6 is one of the most important mediators in the regulation of the immune response, hematopoiesis, and inflammation [1]. It plays roles in chronic inflammation, autoimmune diseases, and cancer. In addition, it has been reported to be involved in the cytokine storm triggered by the corona virus disease 2019 (COVID-19) [2].
 

IL-6 exerts its action by first binding to the IL-6 receptor (IL-6R). The complex of IL-6 and IL-6R associates with the signal-transducing membrane protein gp130, thereby inducing its dimerization (figure 1). This leads to the downstream activation by phosphorylation of certain tyrosine kinases within the Janus family (JAK1, JAK2, and Tyk2). Subsequently, activated JAKs induce phosphorylation, dimerization and translocation of the signal transducer and activator of transcription 3 (STAT3) to the nucleus where it binds enhancer elements of IL-6 inducible genes [3]. Considering the various properties of this pleiotropic cytokine, the IL-6–STAT3 signaling axis is now considered a major target for the treatment of inflammatory diseases [2].

References:

1. Kang S. et al., 2015. Therapeutic uses of anti-interleukin-6 receptor antibody. Int. Immunol. 27: 21-9.
2. Hirano T., 2021. IL-6 in inflammation, autoimmunity and cancer. Int Immunol. 1;33(3):127-148.
3. Murray P., 2007. The JAK-STAT Signaling Pathway: Input and Output Integration. J. Immunol. 178:2623-9.