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Questions? Please consult our answers to frequently asked questions at http://www.ebioscience.com/faq. DescriptionThe TN3-19.12 monoclonal antibody reacts with mouse, rat, and rabbit tumor necrosis factor-alpha (TNF-α), but not with human TNF-α. The TN3-19.12 antibody is a neutralizing antibody. Mouse TNF-α is a 17 kDa factor produced by macrophages, monocytes, neutrophils, CD4+ T cells and NK cells. A 26 kDa form of TNF-α is expressed as a membrane-bound molecule. TNF-α is cytolytic and plays an important role in immune regulation. Dimers and trimers of TNF-α have been observed. Applications ReportedFor research use only, not for diagnostic or therapeutic use. 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 # 16-7325), is recommended. Applications TestedThe TN3-19.12 antibody has been tested as the capture antibody in a sandwich ELISA for analysis of mouse or rat TNF-α in combination with the biotinylated anti-mouse/rat TNF-α polyclonal antibody (13-7341) for detection and recombinant mouse TNF-α (14-8321) or rat TNF-α as the standard. [For a rat TNF-α ELISA standard, please consult eBioscience Technical Support. Product 14-8320 is a bioactive protein, but is not recommended for rat TNF-α 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 recombinant standard over the range of 2000 pg/ml - 15 pg/ml (mouse TNF-α) or 5000 – 30 pg/ml (rat TNF-α) should be included in each ELISA plate. Recognition of mouse TNF-α by this assay is not interfered with by 1000x excess soluble TNF-α receptor type I or II. The Functional Grade purified TN3-19.12 antibody has been tested for neutralization of TNF-α 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/million cells. 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 million cells. References
Sheehan KC, Ruddle NH, Schreiber RD. 1989. Generation and characterization of hamster monoclonal antibodies that neutralize murine tumor necrosis factors. J Immunol. 142: 3884-93. Rabinovici R, Bugelski PJ, Esser KM, Hillegass LM, Griswold DE, Vernick J, Feuerstein G. 1993. Tumor necrosis factor-alpha mediates endotoxin-induced lung injury in platelet activating factor-primed rats. Macol Exp Ther. 267: 1550-57. Merrick BA, He CY, Craig WA, Clark GC, Corsini E, Rosenthal GJ, Mansfield BK, Selkirk JK. 1992. Two dimensional gel electrophoresis of cellular and secreted proteins from rat alveolar macrophages after lipopolysaccharide treatment. Appl Theor Electrophor. 2: 177-87. Takahashi S, Kapas L, Fang J, Krueger JM. 1995. An anti-tumor necrosis factor antibody suppresses sleep in rats and rabbits. Brain Res. 690: 241-44. 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. Finkelman, F.D., and S.C. Morris. 1999. Development of an assay to measure in vivo cytokine production in the mouse. Int. Immunology. 11: 1811-1818. Related ProductsCat. 00-0000 eBioscience 10X ELISA Coating Buffer Cat. 44-2404 Nunc MaxiSorp™ flat-bottom 96 well plate Cat. 18-4100 Avidin Horse Radish Peroxidase (AV-HRP) Cat. 00-4202 eBioscience ELISA Diluent Solution, 5X Cat. 00-4203 Super AquaBlue ELISA Substrate Cat. 14-4888 Affinity Purified Armenian Hamster IgG Isotype Control (clone eBio299Arm) Cat. 11-7321 FITC anti-mouse TNFa (Tumor Necrosis Factor alpha, TNF-alpha, TNF-a) (clone MP6-XT22) Cat. 12-7321 PE anti-mouse TNFa (Tumor Necrosis Factor alpha, TNF-alpha, TNF-a) (clone MP6-XT22) Cat. 14-7321 Affinity Purified anti-mouse TNFa (Tumor Necrosis Factor alpha, TNF-alpha, TNF-a) (clone MP6-XT22) Cat. 17-7321 APC anti-mouse TNFa (Tumor Necrosis Factor alpha, TNF-alpha, TNF-a) (clone MP6-XT22) Cat. 19-7321 Cy5 anti-mouse TNFa (Tumor Necrosis Factor alpha, TNF-alpha, TNF-a) (clone MP6-XT22) Cat. 16-7322 Functional Grade Purified anti-mouse TNFa (Tumor Necrosis Factor alpha, TNF-alpha, TNF-a) (clone MP6-XT3) Cat. 88-7324 Mouse TNFa (Tumor Necrosis Factor alpha, TNF-alpha, TNF-a) ELISA Ready-SET-Go! Cat. 14-7325 Affinity Purified anti-mouse TNFa (Tumor Necrosis Factor alpha, TNF-alpha, TNF-a) (clone 1F3F3D4) Cat. 13-7326 Biotin anti-mouse TNFa (Tumor Necrosis Factor alpha, TNF-alpha, TNF-a) Cocktail (clone XT3/XT22) Cat. 13-7341 Biotin anti-mouse/rat TNFa (Tumor Necrosis Factor alpha, TNF-alpha, TNF-a) polyclonal (clone Polyclonal) Cat. 00-8222 eBioscience IC Fixation Buffer Cat. 14-8320 Recombinant Rat TNFa (Tumor Necrosis Factor alpha, TNF-alpha, TNF-a) Cat. 14-8321 Recombinant Mouse TNFa (Tumor Necrosis Factor alpha, TNF-alpha, TNF-a) Cat. 00-8333 eBioscience Permeabilization Buffer(10X) Cat. 88-8823 eBioscience Fixation & Permeabilization |
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