Intra-Assay Precision | Inter-Assay Precision | |||||
---|---|---|---|---|---|---|
Sample | 1 | 2 | 3 | 1 | 2 | 3 |
n | 20 | 20 | 20 | 20 | 20 | 20 |
Mean | 54.6 | 116 | 405 | 52.6 | 138 | 373 |
Standard Deviation | 1.5 | 4.51 | 12.5 | 4.61 | 8.49 | 29.7 |
CV% | 2.7 | 3.9 | 3.1 | 8.8 | 6.2 | 8 |
Intra-Assay Precision | Inter-Assay Precision | |||||
---|---|---|---|---|---|---|
Sample | 1 | 2 | 3 | 1 | 2 | 3 |
n | 20 | 20 | 20 | 20 | 20 | 20 |
Mean | 50.5 | 134 | 375 | 53 | 146 | 388 |
Standard Deviation | 1.91 | 4.95 | 11.5 | 3.69 | 8.42 | 29.7 |
CV% | 3.8 | 3.7 | 3.1 | 7 | 5.8 | 7.7 |
The recovery of mouse 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=6) | 104 | 94-111 |
EDTA Plasma (n=4) | 95 | 85-103 |
Heparin Plasma (n=4) | 92 | 86-98 |
Serum (n=4) | 94 | 88-106 |
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.
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