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, ELISPOT, IVCCA, 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 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 alpha) or 5000 - 30 pg/mL (rat TNF alpha) should be included in each ELISA plate. Recognition of mouse TNF alpha 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. The fluorochrome-conjugated TN3-19.12 antibody is offered in 2 formats: - µg size: has been tested by intracellular staining and flow cytometric analysis and can be used at less than or equal to 0.25 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. - test size: has been pre-titrated and tested by intracellular staining and flow cytometric analysis and can be used at 20 µL per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC.Filtration: 0.2 µm post-manufacturing filtered. 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.
|ELISA, Flow Cytometry, Functional Assay, Neutralization|
|PBS with 0.09% sodium azide; pH 7.2|
For Research Use Only.
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