Product Information

Contents: In Vivo Cytokine Capture Assay (IVCCA) In Vivo Capture Antibody / Functional Grade Biotin anti-mouse TNF-α
Catalog Number: 36-7423
Sizes: 500 ug
Formulation: Phosphate buffer pH 7.2, 150 mM NaCl, No NaN3. Endotoxin level is less than 0.05 ng/ug antibody, as determined by the LAL assay.
Storage Conditions: Store at 4°C. Do not freeze. Keep contents sterile.
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 Functional Grade Biotin anti-mouse TNF-α 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 this reference: 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-α in combination with the purified polyclonal anti-mouse TNF-α antibody (14-7341) for capture and recombinant mouse TNF-α (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.

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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