Tumor necrosis factor alpha (TNF-alpha), also known as cachectin; and tumor necrosis factor beta (TNF-alpha), also known as lymphotoxin, are two closely related proteins (approximately 34%aa sequence identity) that bind to the same cell surface receptors and show many common biological functions. TNF-alpha and -beta play a critical role in normal host resistance to infection and to the growth of malignant tumors, serving as immunostimulants and as mediators of the inflammatory response. Over-production of TNFs, however, has been implicated as playing a role in a number of pathological conditions, including cachexia, septic shock, and autoimmune disorders. TNF-alpha is produced by activated macrophages and other cell types including T and B cells, NK cells, LAK cells, astrocytes, endothelial cells, smooth muscle cells and some tumor cells. Mouse TNF-alpha cDNA encodes a 235aa residue type II membrane protein.
Sample Type:Cell culture supernatants, serum, plasma, and tissue
Intended Use:The Mouse TNF-alpha BioAssay™ ELISA Kit is an enzyme-linked immunosorbent assay for the quantitative detection of Mouse TNF-alpha concentrations in cell culture supernatants, serum, plasma, and tissue.
Sensitivity:7pg/ml
Range:31.25-2000pg/ml
Specificity:Recognizes mouse TNF-alpha.
Sample Volume:100ul/well
Test Principle:This assay employs the Sandwich immunoassay technique. An anti-m TNF-alpha monoclonal coating antibody is adsorbed onto microwells. TNF-alpha present in the sample or standard binds to antibodies adsorbed to the microwells. Following incubation unbound sample or standard are removed during a wash step. a Biotinylated anti-m TNF-alpha antibody is added and binds to TNF-alpha captured by the first antibody. Following incubation unbound Biotinylated anti-m TNF-alpha antibody is removed during a wash step. A Streptavidin-HRP is added and binds to Biotinylated anti-m TNF-alpha antibody. Following incubation unbound Streptavidin-HRP is removed during a wash step. A colored product is formed in proportion to the amount of TNF-alpha present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven TNF-alpha standard dilutions and TNF-alpha sample concentration determined.
Storage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
