The NWLSS™ Rat TNF-α ELISA kit is intended to be used for the in vitro quantitative determination of Tumor Necrosis factor Alpha (TNF-α) in rat serum, plasma, cell lysates and cell culture supernatants. The assay will recognize native and recombinant rat TNF-α.
Tumor necrosis factor alpha (TNF-α) is an important cytokine in the innate immune response. It is primarily produced by macrophages but is also produced by other cell types including T-Lymphocytes, Natural Killer (NK), endothelial and neuronal cells. TNF-α is initially produced as a biologically active 26 kDa transmembrane protein, which is subsequently cleaved, principally by TNF-α-converting enzyme (TACE), to release a 17 kDa free protein. Free TNF-α is a homotrimer that acts on TNF-α receptors 1 and 2 (TNFR1 and TNFR2) expressed on many different cell types. Both free and transmembrane TNF-α are capable of mediating host responses in acute and chronic inflammatory conditions and have also been shown to be involved in apoptotic cell death, cellular proliferation, differentiation and tumorigenesis. TNF-α has been shown to be a key player in the inflammatory response associated with autoimmune disorders such as rheumatoid arthritis, inflammatory bowel disease (IBD), psoriasis, asthma and others. As such TNF-α blockade has become a common strategy in treating several types of chronic inflammatory disease.
The NWLSS™ Rat TNF-α ELISA is a sandwich format Enzyme-Linked Immunosorbent Assay (ELISA). The microtiter plate provided in this kit has been pre-coated with a monoclonal antibody specific to rat TNF-α. Samples are pipetted into these wells. Non-bound TNF-α and other components of the sample are removed by washing, after which a biotinylated antibody specific to rat TNF-α is added. In order to quantitatively determine the amount of TNF-α present in the sample, Streptavidin Horseradish Peroxidase (HRP) conjugate is added to each microplate well. After another wash step, TMB-substrate solution is added to each well. Finally, a sulfuric acid stop solution is added and the resulting yellow colored product is measured at 450nm. The amount of TNF-α in the sample can be determined by direct comparison with the standard curve generated in the assay.
View Assay Protocol
1 X 96 well ELISA presented as 12 X 8 well (6 X 16 well) strips in frame.
For quantitative detection of rat TNF-α.
Rat serum, plasma, cell lysates and cell culture supernatants.
|Storage and Stability:||
9 months from date of manufacture
1 Foil Pouch 96 well microplate precoated with anti-Rat TNF-α.|
2 bottles 20X Concentrated Wash Buffer (25 mL)
1 vial rRat TNF-α Standard (lyophilized) (1 Vial)
1 bottle Standard/Sample Dilution Buffer (25mL)
1 vial Secondary Antibody (Lyophilized) (1 Vial) (Biotinylated Anti-Rat TNF-α)
1 vial 100X Streptavidin-HRP Conjugate (150 µL)
1 bottle Reagent Dilution Buffer (25mL)
1 bottle TMB Substrate (15 mL)
1 bottle Stop Solution (1 N Sulfuric Acid, H2SO4) (15 mL)
2 Adhesive Plate Covers (2)