Product Information

Contents: Functional Grade Purified anti-mouse/rat TNF-alpha (TNF-α)
Catalog Number: 16-7423
Sizes: 50 ug, 500 ug
Formulation: Phosphate buffer pH 7.2,
150 mM NaCl, No NaN₃
Storage Conditions: Store at 4°C.
Avoid repeated freeze/thaw cycles.
KEEP CONTENTS STERILE.
Endotoxin Level: Less than 0.001 ng/ug antibody, as determined by the LAL assay.
Clone: TN3-19
Isotype: Armenian Hamster IgG

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
11-7423	Fluorescein (FITC) anti-mouse/rat TNFa (Tumor Necrosis Factor alpha, TNF-alpha, TNF-a)	488	518	IC Flow
12-7423	Phycoerythrin (PE) anti-mouse/rat TNFa (Tumor Necrosis Factor alpha, TNF-alpha, TNF-a)	488	575	IC Flow
14-7423	Affinity Purified anti-mouse/rat TNFa (Tumor Necrosis Factor alpha, TNF-alpha, TNF-a)	N/A	N/A	ELISA cap IC Flow NU
16-7423	Functional Grade* Purified anti-mouse/rat TNFa (Tumor Necrosis Factor alpha, TNF-alpha, TNF-a)	N/A	N/A	NU
25-7423	Phycoerythrin-Cy7 (PE-Cy7) anti-mouse/rat TNFa (Tumor Necrosis Factor alpha, TNF-alpha, TNF-a)	488	760	ELISA IC Flow NU
36-7423	In Vivo Cytokine Capture Assay (IVCCA) In Vivo Capture Antibody / Functional Grade* Biotin anti-mouse TNFa (Tumor Necrosis Factor-alpha, TNF-a)	N/A	N/A	IVCCA
*Functional Grade™ (FG™) Purified: Azide-free, sterile-filtered, and endotoxin < 0.001 ng/µg (unless otherwise noted). *Functional Grade™ (FG™) Biotin: Azide-free, sterile-filtered, and endotoxin < 0.05 ng/µg. Purified: Contains azide, not sterile-filtered, and not endotoxin tested.

Questions? Please consult our answers to frequently asked questions at http://www.ebioscience.com/faq.

Description

The 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 Reported

For 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, 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 Purified TN3-19.12 antibody has been tested by LAL assay to verify low endotoxin level and has been tested for neutralization of TNF-α bioactivity and for ELISA capture.

The TN3-19.12 antibody at 0.150 ug/ml has been found to inhibit by 50% the biological effects of 1.0 ng/ml mouse TNF-α 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-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-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-a receptor type I or II. The Functional Grade purified TN3-19.12 antibody has been tested for neutralization of TNF-a bioactivity.

The fluorochrome-conjugated TN3-19.12 antibody is offered in 2 formats:
- ug size: has been tested by intracellular staining and flow cytometric analysis and can be used at less than or equal to 0.25 ug/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 ul 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.

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