Tumor necrosis factor alpha (TNF-α), also known as cachectin and TNFSF2, is the prototypic ligand of the TNF superfamily. It is a pleiotropic molecule that plays a central role in inflammation, apoptosis, and immune system development. TNF-α is produced by a wide variety of immune and epithelial cell types. Human TNF-α consists of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 177 aa extracellular domain (ECD). Within the ECD, human TNF-α shares 97% aa sequence identity with rhesus and 71% - 92% with bovine, canine, cotton rat, equine, feline, mouse, porcine, and rat TNF-α. The 26 kDa type 2 transmembrane protein is assembled intracellularly to form a noncovalently linked homotrimer. Ligation of this complex induces reverse signaling that promotes lymphocyte costimulation but diminishes monocyte responsiveness.
Cleavage of membrane bound TNF-α by TACE/ADAM17 releases a 55 kDa soluble trimeric form of TNF-α. TNF-α trimers bind the ubiquitous TNF RI and the hematopoietic cell-restricted TNF RII, both of which are also expressed as homotrimers. TNF-α regulates lymphoid tissue development through control of apoptosis. It also promotes inflammatory responses by inducing the activation of vascular endothelial cells and macrophages. TNF-α is a key cytokine in the development of several inflammatory disorders. It contributes to the development of type 2 diabetes through its effects on insulin resistance and fatty acid metabolism.
原厂资料：
Product Summary:
The QuantiGlo Human TNF-alpha Chemiluminscent Immunoassay is a 5 hour solid phase ELISA designed to measure TNF-alpha in cell culture supernates, serum, and plasma. It contains E. coli-derived recombinant human TNF-alpha and antibodies raised against the recombinant factor. It has been shown to accurately quantitate recombinant human TNF-alpha. Results obtained using naturally occurring TNF-alpha samples showed linear curves that were parallel to the standard curves obtained using the QuantiGlo kit standards. These results indicate that this kit can be used to determine relative mass values for natural TNF-alpha.
Precision:
Intra-Assay Precision (Precision within an assay)Four samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays)Four samples of known concentration were tested in separate assays to assess inter-assay precision.
The recovery of TNF-alpha spiked to three different levels throughout the range of the assay in various matrices was evaluated.
Sample Type
Average % Recovery
Range %
Cell Culture Media (n=4)
107
97-115
Serum (n=4)
101
87-113
EDTA Plasma (n=4)
100
86-112
Heparin Plasma (n=4)
100
83-114
Citrate Plasma (n=4)
100
91-109
Linearity:
To assess the linearity of the assay, samples spiked with high concentrations of TNF-alpha in various matrices were diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
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