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 Quantikine Porcine TNF-alpha Immunoassay is a 4.5 hour solid phase ELISA designed to measure porcine TNF-alpha in cell culture supernates, serum, and plasma samples. It contains E. coli-expressed recombinant porcine TNF-alpha and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately measure recombinant porcine TNF-alpha. Results obtained using natural porcine TNF-alpha show dose response curves that are parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural porcine TNF-alpha.
Precision:
Intra-Assay Precision (Precision within an assay)Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays)Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision
Inter-Assay Precision
Sample
1
2
3
1
2
3
n
20
20
20
20
20
20
Mean
104
358
880
106
348
866
Standard Deviation
7.2
13
36
9.7
32
72
CV%
6.9
3.6
4.1
9.2
9.2
8.3
Cell Culture Supernates
Intra-Assay Precision
Inter-Assay Precision
Sample
1
2
3
1
2
3
n
20
20
20
20
20
20
Mean
79
260
646
88
279
679
Standard Deviation
4.9
9.2
27
8.8
24
44
CV%
6.2
3.5
4.2
10
8.6
6.5
Recovery:
The recovery of porcine TNF-alpha spiked to three levels throughout the range of the assay in various matrices was evaluated.
Sample Type
Average % Recovery
Range %
Serum (n=6)
105
91-120
Heparin Plasma (n=4)
91
82-99
Cell Culture Supernates (n=6)
99
91-105
EDTA Plasma (n=6)
98
83-111
Linearity:
To assess the linearity of the assay, six samples containing and/or spiked with various concentrations of porcine TNF-alpha in each matrix were diluted with the appropriate Calibrator Diluent and then assayed.
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