Name
TNF-alpha, Human, ELISA kit
Catalog nr
HK307
(lot number and expiry date are indicated on the label)
Short description
Tumor necrosis factor alpha (TNF-alpha), a homotrimeric 17 KD protein, is a potent mediator of inflammatory and metabolic functions. TNF-alpha was originally detected as a highly cytotoxic cytokine for tumor cells, it causes tumor necrosis in vivo and shows cytolytic activity against tumor cells in vitro. Furthermore, TNF-alpha has been implied as central mediator in shock induced by gram neg...
Size
2 x 96 det.
Application
Manual
Description
Tumor necrosis factor alpha (TNF-alpha), a homotrimeric 17 KD protein, is a potent mediator of inflammatory and metabolic functions. TNF-alpha was originally detected as a highly cytotoxic cytokine for tumor cells, it causes tumor necrosis in vivo and shows cytolytic activity against tumor cells in vitro. Furthermore, TNF-alpha has been implied as central mediator in shock induced by gram negative micro-organisms.
The cytokine TNF is found to be a central mediator in many inflammatory and immunological processes: it can be induced by various products of micro-organisms and by various cytokines but it also induces on its turn the production of many cytokines.
Signal transduction occurs via two types of TNF-receptors, the p55 (RI) and p75 (RII) TNF-receptors. The receptors differ strongly in their intra-cellular signaling pathways.
The TNF-alpha trimer interacts with either of the two types of TNF-R leading to receptor cross-linking.
Application
The human TNF-alpha ELISA has been developed for the quantitative measurement of natural and recombinant human TNF-alpha in serum, plasma, and cell culture medium. In serum or plasma samples human TNF-alpha can be measured accurately if serum or plasma samples are diluted at least 4 times. Most reliable results are obtained if EDTA plasma is used.
Features
- Minimum concentration which can be measured is 10 pg/ml human TNF-alpha.
- Measurable concentration range of 8-2,000 pg/ml.
- Working volume of 100 µl/well.
Typical standard curve
Principle
- The human TNF-alpha ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle with a working time of 3½ hours.
- The efficient format of 2 plates with twelve disposable 8-well strips allows free choice of batch size for the assay.
- Samples and standards are captured by a solid bound specific antibody.
- Biotinylated tracer antibody will bind to captured human TNF-alpha.
- Streptavidin-peroxidase conjugate will bind to the biotinylated tracer antibody.
- Streptavidin-peroxidase conjugate will react with the substrate, tetramethylbenzidine (TMB).
- The enzyme reaction is stopped by the addition of citric acid.
- The absorbance at 450 nm is measured with a spectrophotometer. A standard curve is obtained by plotting the absorbance (linear) versus the corresponding concentrations of the human TNF-alpha standards (log).
- The human TNF-alpha concentration of samples, which are run concurrently with the standards, can be determined from the standard curve.
Storage and stability
Product should be stored at 4°C. Under recommended storage conditions, product is stable for at least six months. After reconstitution the reagents are stable for 1 month if stored at 2-8°C, except for the standard. The standard is stable for 15 minutes after reconstitution. For longer stability we recommend to store aliquots at -20°C.
References
- Molnár, L et al; Detection of TNFalpha expression in the bone marrow and determination of TNFalpha production of peripheral blood mononuclear cells in myelodysplastic syndrome. Pathol Oncol Res 2000, 6: 18
- Younis, A et al; Staphylococcus aureus leucocidin, a virulence factor in bovine mastitis. J Dairy Research 2005, 72: 188
Precautions
For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and Federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result with the use of or derivation of this product.
Also available
References
- Molnár, L et al; Detection of TNFalpha expression in the bone marrow and determination of TNFalpha production of peripheral blood mononuclear cells in myelodysplastic syndrome. Pathol Oncol Res 2000, 6: 18
- Younis, A et al; Staphylococcus aureus leucocidin, a virulence factor in bovine mastitis. J Dairy Research 2005, 72: 188
MSDS
| Related products | Cat # | |
|---|---|---|
| IFN-gamma, Human, ELISA kit (medium & serum) | HK309 | |
| sTNF-RII, Human, ELISA kit | HK302 | |
| sTNF-RI, Human, ELISA kit | HK301 | |