CellaQuantTM Mouse TNF-α (Tumor Necrosis Factor Alpha) ELISA Kit

Tumor Necrosis Factor-alpha (TNF-α), also known as cachectin or TNFSF1A, is the prototypic ligand of the Tumor Necrosis Factor superfamily. As a pleiotropic cytokine, TNF-α plays a central role in immunity, inflammation, and metabolic homeostasis. Its functions are primarily mediated by two receptors, TNFR1 and TNFR2, which activate signaling pathways such as NF-κB and MAPK to regulate a broad spectrum of cellular responses. In terms of immunity, TNF-α activates innate immune cells including macrophages and neutrophils, promotes the release of inflammatory cytokines such as IL-1 and IL-6, and modulates the activation and differentiation of T and B lymphocytes. In tissue homeostasis, TNF-α can induce apoptosis in target cells while also promoting cell survival via NF-κB signaling; under certain conditions, it also contributes to the induction of necroptosis. Metabolically, TNF-α inhibits lipoprotein lipase activity, leading to cachexia, and is involved in the development of insulin resistance. Dysregulation of TNF-α is closely associated with a variety of diseases, including autoimmune disorders such as rheumatoid arthritis and Crohn's disease, asthma, neuropathic pain, obesity and type 2 diabetes, septic shock, and the progression of various cancers. Human TNF-α is initially synthesized as a 26 kDa type II transmembrane protein, consisting of an intracellular domain of 35 amino acids, a transmembrane segment of 21 amino acids, and an extracellular domain of 177 amino acids. It is produced by a wide range of immune cells (such as macrophages and T cells), epithelial cells, endothelial cells, and certain tumor cells.

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Mouse TNF-α. Samples (or Standards &Quality Control Material) are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Mouse TNF-α and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Mouse TNF-α, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450±2 nm. The OD value is proportional to the concentration of Mouse TNF-α. You can calculate the concentration of Mouse TNF-α in the samples by comparing the OD of the samples to the standard curve.

This ELISA kit applies to the in vitro quantitative determination of Mouse TNF-α concentrations in Cell culture supernatant, Cell lysates or other biological fluids.