Mouse TNF-alpha Quantikine HS ELISA Kit

  
  • Assay Length
    4.0 hours
  • Sample Type & Volume Required Per Well
    Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL)
  • Sensitivity
    0.295 pg/mL
  • Assay Range
    0.8 - 50 pg/mL (Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant mouse TNF-alpha.
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Product Summary
The Quantikine HS Mouse TNF-alpha Immunoassay is a 4.0 hour solid phase ELISA designed to measure mouse TNF-alpha levels in serum and plasma. It contains E. coli-expressed recombinant mouse TNF-alpha and antibodies raised against the recombinant factor and has been shown to accurately quantitate recombinant mouse TNF-alpha. Results obtained using natural TNF-alpha showed linear curves that were parallel to the standard curves obtained using the Quantikine HS kit standards. These results indicate that the Quantikine HS Immunoassay kit can be used to determine relative mass values for natural mouse TNF-alpha.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean6.6315.129.26.2915.228.5
Standard Deviation0.2570.6951.490.5521.181.94
CV%3.94.65.18.87.86.8

Recovery

The recovery of mouse TNF-alpha spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
EDTA Plasma (n=4) 91 81-104
Heparin Plasma (n=4) 90 79-106
Serum (n=4) 86 77-99
Linearity
To assess the linearity of the assay, samples spiked with high concentrations of mouse TNF-alpha were serially diluted with calibrator diluent to produce samples with values within the dynamic range of the assay.
Mouse TNF-alpha Quantikine HS ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: TNF-alpha

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.

    • Long Name
      Tumor Necrosis Factor alpha
    • Entrez Gene IDs
      7124 (Human); 21926 (Mouse); 24835 (Rat); 397086 (Porcine); 280943 (Bovine); 403922 (Canine); 100033834 (Equine); 493755 (Feline); 100009088 (Rabbit);
    • Alternate Names
      APC1 protein; Cachectin; Cachetin; DIF; TNF; TNF, monocyte-derived; tnfa; tnf-a; TNFalpha; TNF-alpha; TNF-alphacachectin; TNFATNF, macrophage-derived; TNFSF1A; TNFSF2; TNFSF2TNF superfamily, member 2; tumor necrosis factor (TNF superfamily, member 2); tumor necrosis factor alpha; Tumor necrosis factor ligand superfamily member 2; tumor necrosis factor; tumor necrosis factor-alpha;
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