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 Rhesus Macaque TNF-alpha Immunoassay is a 4.5 hour solid phase ELISA designed to measure rhesus macaque TNF-alpha levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant rhesus macaque TNF-alpha and antibodies raised against recombinant rhesus macaque TNF-alpha. This immunoassay has been shown to quantitate the recombinant rhesus macaque TNF-alpha accurately. Results obtained using natural rhesus macaque TNF-alpha showed dose curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural rhesus macaque 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.
Cell Culture Supernates, Serum, EDTA Plasma
Intra-Assay Precision
Inter-Assay Precision
Sample
1
2
3
1
2
3
n
20
20
20
20
20
20
Mean
61
123
492
56
111
465
Standard Deviation
3.6
7.4
35.7
5.6
7.2
41.8
CV%
5.9
6
7.3
10
6.5
9
Recovery:
The recovery of rhesus macaque TNF-alpha spiked to three levels throughout the range of the assay in various matrices was evaluated.
Sample Type
Average % Recovery
Range %
Cell Culture Supernates (n=4)
110
104-120
Serum (n=4)
98
90-109
EDTA Plasma (n=3)
94
89-99
Linearity:
To assess the linearity of the assay, samples containing and/or spiked with various concentrations of rhesus macaque TNF-alpha in each matrix were diluted with Calibrator \line Diluent and then assayed.
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