Capture Antibody: Pre-titrated, purified antibody
Detection Antibody: Pre-titrated, biotin-conjugated antibody
Standard: Recombinant cytokine for generating standard curve and calibrating samples
ELISA/ELISPOT Coating Buffer Powder: This uncoated ELISA kit with platest may contain ELISA/ELISPOT Coating Buffer Powder (Reconstitute to 1L with dH20 and filter (0.22 µM)) or 10X PBS ELISA Coating Buffer (Dilute 1 part 10X Buffer into 9 parts dH20).
Assay Diluent: 5X concentrated
Detection enzyme: Pre-titrated Avidin-HRP
Substrate Solution: Tetramethylbenzidine (TMB) Substrate Solution
Certificate of Analysis: Lot-specific instructions for dilution of antibodies and standards
96 Well Plate: Corning Costar 9018 (included with product Cat. Nos. ending in suffixes -22, -44, -76, -86)

C-domain 1, C-domain 2, Cachectin, ICD1, ICD2, Intracellular domain 1, Intracellular domain 2, N-terminal fragment, NTF, TNF-a, TNF-alpha, Tumor necrosis factor, tumor necrosis factor (TNF superfamily, member 2), tumor necrosis factor alpha, Tumor necrosis factor ligand superfamily member 2, membrane form, soluble form

The Rat Tumor Necrosis Factor Alpha (TNF-α) Uncoated ELISA Kit contains pre-matched antibody pairs, plates and reagents for performing quantitative enzyme linked immunosorbent assays (ELISA) to detect and quantify protein levels of rat TNF-α. Wash Buffer and Stop Solution are needed to complete the ELISA reaction and are sold separately.

Principle of the method

ELISAs are designed to measure the amount of the target bound between a matched antibody pair. A target-specific antibody is coated to the bottom of the wells of a microplate, which is an overnight process. Samples, standards, or controls are then added into these wells and bind to the immobilized (capture) antibody. A sandwich is formed by the addition of the second (detector) antibody, a substrate solution is added that reacts with the enzyme-antibody-target complex to produce measurable signal. The intensity of this signal is directly proportional to the concentration of target present in the original specimen.

TNF alpha is a multifunctional proinflammatory cytokine that belongs to the tumor necrosis factor (TNF) superfamily. This cytokine is mainly secreted by macrophage and bind to its receptors, TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. TNF alpha is involved in the regulation of immune cells, cell proliferation, differentiation, apoptosis, lipid metabolism, and coagulation. TNF alpha exists as a multimer of two, three, or five noncovalently linked units, but shows a single 17 kDa band following SDS-PAGE under non-reducing conditions. Knockout studies in mice also suggested the neuroprotective function of TNF alpha, and has been observed to causes tumor necrosis when injected into tumor-bearing mice. Other functions of TNF-alpha include its role in the immune response to bacterial, viral, parasitic and certain fungal infections, as well as its role in the necrosis of specific tumors. TNF alpha causes cytolysis or cytostasis of certain transformed cells, being synergistic with interferon-gamma in its cytotoxicity. This cytokine has been implicated in a variety of diseases, including autoimmune diseases, insulin resistance, and cancer.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.