Product Information

Contents: Phycoerythrin-Cy7 (PE-Cy7) anti-mouse/rat TNF-α
Catalog Number: 25-7423
Sizes: 25 tests, 100 ug
Formulation: Phosphate buffered saline pH 7.2, less than or equal to 0.09% NaN₃, protein stabilizer
Storage Conditions: Store at 4°C.
DO NOT FREEZE.
LIGHT-SENSITIVE MATERIAL.
This Cy7 tandem dye is sensitive to photo-induced oxidation. Protect this vial from light during storage, handling & experimental procedures.
This product has been improved, resulting in brighter staining with less compensation required. For more information see PE-Cy7.
Clone: TN3-19
Isotype: Armenian Hamster IgG

Mouse splenocytes were stimulated with ConA for 2 days, followed by IL-2 and IL-4 for 3 days, and restimulated with immobilized anti-CD3 and soluble anti-CD28 in the presence of Brefeldin A for 5 hours. The cells were surface stained with APC anti-mouse CD4 (GK1.5) (cat. 17-0041) and intracellularly stained with PE-Cy7 anti-mouse TNF-a (TN3-19.12).

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. This TN3-19 antibody has been reported for use in intracellular staining followed by flow cytometric analysis, cytokine neutralization, and ELISA.

This tandem dye is sensitive to photo-induced oxidation. Protect this reagent from light during storage and all handling steps. When staining cells with PE-Cy7-conjugated antibodies, always protect samples from light.

Applications Tested

This TN3-19 antibody is offered in 2 formats:

- μg size: has been tested by intracellular staining and flow cytometric analysis of ConA-stimulated mouse splenocytes. This can be used at less than or equal to 1 μg per 100 μl blood (or per 1 million cells in 100 μl total staining volume). 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 of ConA-stimulated mouse splenocytes. This can be used at 20 μl (1 μg) per 100 μl blood (or per 1 million cells in 100 μl total staining volume). The pre-titrated test size contains BSA and/or gelatin for protein stabilization.

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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FOR NON-COMMERCIAL RESEARCH USE ONLY. NOT FOR THERAPEUTIC OR IN VIVO APLICATIONS. OTHER USE NEEDS LICENSE FROM AMERSHAM PHARMACIA BIOTECH INC. UNDER U.S. PATENT Nos. 4,981,977 AND 5,268,486 AND OTHER PATENTS PENDING.

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