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 Functional Grade Biotin anti-mouse TNF alpha antibody (clone TN3-19) has been reported for use as the in vivo capture antibody for the In Vivo Cytokine Capture Assay (IVCCA). The IVCCA facilitates measurement of cytokines in serum by increasing their in vivo half lives dramatically. This increases the sensitivity of measurement of in vivo cytokine production 30- to 1000- fold.
For detailed methodology in this application, please refer to Finkelman, F., S. Morris, T. Orekhova, and D. Sehy. 2003. The In Vivo Cytokine Capture Assay for measurement of cytokine production in the mouse. In Current Protocols in Immunology. Unit 6.28. J. Coligan, A. Kruisbeek, D. Margulies, E. Shevach, and W. Strober, eds. John Wiley and Sons, New York 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 # 16-7325), is recommended.
Applications Tested: The Functional Grade Biotin TN3-19 antibody has been tested by LAL assay to ensure it is free of endotoxin, and has been tested as the detection antibody in a sandwich ELISA for analysis of mouse TNF alpha in combination with the purified polyclonal anti-mouse TNF alpha antibody (14-7341) for capture and recombinant mouse TNF alpha (14-8321) as the standard.A suitable range of concentrations of the FG Biotin TN3-19 antibody for ELISA detection is 0.5-2 µg/mL. A standard curve consisting of doubling dilutions of the recombinant standard over the range of 2000 pg/mL - 15.0 pg/mL should be included in each ELISA plate.
Endotoxin: Less than 0.05 ng/µg antibody as determined by the LAL assay.
Storage and handling: Use in a sterile environment.
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.
Protein Aliases: C-domain 1; C-domain 2; Cachectin; DADB-70P7.1; ICD1; ICD2; Intracellular domain 1; Intracellular domain 2; N-terminal fragment; NTF; TNF alpha; TNF superfamily; TNF α; TNF-a; TNF-alpha; TNFα; Tumor necrosis factor; tumor necrosis factor (TNF superfamily, member 2); tumor necrosis factor alpha; Tumor necrosis factor ligand superfamily member 2; Tumor necrosis factor, membrane form; Tumor necrosis factor, soluble form; tumor necrosis factor-alpha
Gene Aliases: DIF; RATTNF; Tnf; TNF-a; TNF-alpha; Tnfa; TNFalpha; Tnfsf1a; TNFSF2