TNF alpha Monoclonal Antibody (TN3-19.12), Functional Grade, eBioscience™, Invitrogen™

Description

Description: The TN3-19.12 monoclonal antibody reacts with mouse, rat, and rabbit tumor necrosis factor alpha (TNF alpha), but not with human TNF alpha. The TN3-19.12 antibody is a neutralizing antibody. Mouse TNF alpha is a 17 kDa factor produced by macrophages, monocytes, neutrophils, CD4+ T cells and NK cells. A 26 kDa form of TNF alpha is expressed as a membrane-bound molecule. TNF alpha is cytolytic and plays an important role in immune regulation. Dimers and trimers of TNF alpha have been observed. Applications Reported: The TN3-19.12 antibody has been reported for use in intracellular staining for flow cytometric analysis, cytokine neutralization, and ELISA. Fluorochrome conjugated TN3-19.12 is recommended for use in intracellular staining and flow cytometry. Functional Grade purified TN3-19.12, cat. 16-7423, is recommended for use in functional assays. For ELISPOT capture, the alternative clone, 1F3F3D4 (cat No. 16-7325), is recommended. Applications Tested: The Functional Grade Purified TN3-19.12 antibody has been tested by LAL assay to verify low endotoxin level and has been tested for neutralization of TNF alpha bioactivity and for ELISA capture. The TN3-19.12 antibody at 0.150 μg/mL has been found to inhibit by 50% the biological effects of 1.0 ng/mL mouse TNF alpha in an assay of cytotoxicity of L929 cells.

Detailed information and protocols about cytokine bioassays and in vitro cytokine neutralization using antibodies can be found in the BestProtocols® section. The TN3-19.12 antibody has been tested as the capture antibody in a sandwich ELISA for analysis of mouse or rat TNF alpha in combination with the biotinylated anti-mouse/rat TNF alpha polyclonal antibody (13-7341) for detection and recombit mouse TNF alpha (14-8321) or rat TNF- alpha as the standard. Product 14-8320 is a bioactive protein, but is not recommended for rat TNF alpha ELISA.] A suitable range of concentrations of this antibody for ELISA capture is 1-4 μg/mL. A standard curve consisting of doubling dilutions of the recombit standard over the range of 2000 pg/mL- 15 pg/mL (mouse TNF-a) or 5000 - 30 pg/mL (rat TNF-a) should be included in each ELISA plate. Recognition of mouse TNF a by this assay is not interfered with by 1000x excess soluble TNF alpha receptor type I or II. The Functional Grade purified TN3-19.12 antibody has been tested for neutralization of TNF alpha bioactivity. Endotoxin: Less than 0.001 ng/ug antibody as determined by the LAL assay. Storage and handling: Use in a sterile environment. Filtration: 0.2 μm post-manufacturing filtered. Purity: Greater than 90%, as determined by SDS-PAGE. Endotoxin Level: Less than 0.001 ng/μg antibody, as determined by LAL assay. Aggregation: Less than 10%, as determined by HPLC. 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.