Recombinant human TNF-α

Recombinant Human Tumor Necrosis Factor alpha (with HSA)

Tumor necrosis factor-alpha (TNF-α) is a pleiotropic inflammatory cytokine produced by several types of cells, predominantly activated macrophages. TNF-α plays an important role in the immune response to microbial invasions and in the necrosis of specific tumors. TNF-α exists in two forms; a type II transmembrane protein and a mature soluble protein. The TNF-α transmembrane protein is proteolytically cleaved to yield a soluble protein, which subsequently forms a non-covalently linked homotrimer in solution. The individual subunits of this homotrimer have a relative molecular mass each of 17350 Daltons.

Recombinant human TNF-α is produced in Chinese hamster ovary (CHO) cells transfected with the full-length sequence for the human TNF-α gene. These transfected cells produce and secrete biologically active TNF-α. The recombinant protein is purified by affinity chromatography.

Recombinant human TNF-α can be used to induce NF-κb and AP-1 pathways in HEK-Blue™ TNF-α cells.

Figures

SDS PAGE of the recombinant human (h)TNF-a protein. 1μg of hTNF-α 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 ~17 kDa.

Dose-response in HEK-Blue™ TNF-α cells to recombinant TNF-α cytokine. Cells were stimulated with increasing concentrations of recombinant human TNF-α. 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: P01375

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

Purity: >97%, by SDS-PAGE

Molecular mass: ~ 17 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 TNF-α cells

Contents

• 20 μg of recombinant human TNF-α (with HSA) provided as a white lyophilized powder.
• 1.5 ml endotoxin-free water.

Recombinant human TNF-α is shipped at room temperature.

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