Recombinant human IL-12 protein

Recombinant human interleukin-12 protein (with HSA)

Interleukin-12 (IL-12), also known as IL-12p70, cytotoxic lymphocyte maturation factor (CLMF) and natural killer cell stimulatory factor (NKSF), is primarily produced by dendritic cells and macrophages in response to bacterial pathogens [1,2]. It promotes Th1 responses and is a powerful inducer of interferon gamma (IFN-γ). IL-12 is a heterodimeric 70 kDa cytokine composed of two subunits, p35 and p40.

Recombinant human IL-12 is produced in Chinese hamster ovary (CHO) cells transfected with a fusion gene encoding the p35 and p40 subunits linked by a decapeptide containing two elastin motifs. These transfected cells produce and secrete biologically active IL-12. The recombinant protein is purified by size exclusion chromatography.

The activity of recombinant human IL-12 can be monitored in HEK-Blue™ IL-12 cells which express a STAT4-inducible SEAP reporter gene.

References:

1. Vignali DA. & Kuchroo VK., 2012. IL-12 family cytokines: immunological playmakers. Nat Immunol. 13(8):722-8.
2. Trinchieri G.,1995. Interleukin-12: a proinflammatory cytokine with immunoregulatory functions that bridge innate resistance and antigen-specific adaptive immunity. Annu Rev Immunol. 13:251-76.

Figures

SDS PAGE of the recombinant human (h)IL-12 cytokine. 1 µg of hIL-12 was loaded on a 12% Mini-PROTEAN® TGX Stain-Free™ Precast Gel (Bio-Rad). Detection was performed as per the manufacturer’s instructions. A band was detected at ~70 kDa.

Dose-response in HEK-Blue™ IL-12 cells to recombinant IL-12 cytokine. Cells were stimulated with increasing concentrations of recombinant human IL-12. 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).

Specifications

Source: Mammalian; Chinese hamster ovary (CHO) cells

UniProt ID: P29459

Formulation: Lyophilized from a 0.2 μm filtered solution in PBS (pH 7.4) with human serum albumin (HSA) as a carrier protein

Purity: >90%, by SDS-PAGE

Molecular mass: ~ 70 kDa, by SDS-PAGE

Solubility: 100 μg/ml in water

Endotoxin level: < 0.1 EU/μg protein as determined by the LAL method

Bioactivity: Validated using HEK-Blue IL-12 cells

Contents

• 10 μg of recombinant human IL-12 (with HSA) provided as a white lyophilized powder.
• 1.5 ml endotoxin-free water.

Recombinant human IL-12 is shipped at room temperature.

Upon receipt it should be stored at -20°C.