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Safety Data Sheet (SDS) Brilliant Violet 650 Anti-mouse TNF-alpha Antibody     Product Data Sheet (PDF)    
Brilliant Violet 650™ Anti-mouse TNF-α Antibody
3131665 50 µg $260.00       
Clone: MP6-XT22
Isotype: Rat IgG1, κ
Reactivity: Mouse
Immunogen: E. coli-expressed, recombinant mouse TNF-α
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA).
Preparation: The antibody was purified by affinity chromatography and conjugated with Brilliant Violet 650™ under optimal conditions. The solution is free of unconjugated Brilliant Violet 650™ and unconjugated antibody.
Concentration: 0.2 mg/ml
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application:

ICFC - Quality tested

Application Notes:

ELISA or ELISPOT Detection: The biotinylated MP6-XT22 antibody is useful as a detection antibody for a sandwich ELISA or ELISPOT assay, when used in conjunction with purified 6B8 antibody as the capture antibody.
Flow Cytometry6,11,12: The fluorochrome-labeled MP6-XT22 antibody is useful for intracellular immunofluorescent staining and flow cytometric analysis to identify TNF-α-producing cells within mixed cell populations. 
Neutralization1,5,10: The MP6-XT22 antibody can neutralize the bioactivity of natural or recombinant TNF-α.

Recommended Usage:

Each lot of this antibody is quality control tested by intracellular immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is ≤0.5 µg per million cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.

Brilliant Violet 650™ excites at 405 nm and emits at 645 nm. The bandpass filter 660/20 nm is recommended for detection, although filter optimization may be required depending on other fluorophores used. Be sure to verify that your cytometer configuration and software setup are appropriate for detecting this channel. Refer to your instrument manual or manufacturer for support. Brilliant Violet 650™ is a trademark of Sirigen Group Ltd.

This product is subject to proprietary rights of Sirigen Inc. and is made and sold under license from Sirigen Inc. The purchase of this product conveys to the buyer a non-transferable right to use the purchased product for research purposes only. This product may not be resold or incorporated in any manner into another product for resale. Any use for therapeutics or diagnostics is strictly prohibited. This product is covered by U.S. Patent(s), pending patent applications and foreign equivalents.

Application References:

1. Abrams J, et al. 1992. Immunol. Rev. 127:5. (Neut)
2. Abrams J, et al. 1995. Curr. Prot. Immunol. John Wiley and Sons, New York. Unit 6.20
3. Mo X, et al. 1995. J. Virol. 69:1288.
4. Sarawar S, et al. 1994. J. Immunol. 153:1246.
5. Via C, et al. 2001. J. Immunol. 167:6821. (Neut)
6. Infante-Duarte C, et al. 2000 J. Immunol. 165:6107. (FC)
7. Jacobs M, et al. 2000. Immunology 100:494. (IHC)
8. Marinova-Mutachieva L, et al. 1997. Clin. Exp. Immunol. 107:507. (IHC)
9. Williams RO, et al. 2000. J. Immunol. 165:7240. (IHC)
10. Scanga CA, et al. 1999. Infect. Immun. 67:4531. (Neut)
11. Akilov OE, et al. 2007. J. Leukoc. Biol. 2007;10.1189/jlb.0706439. (FC)
12. Lawson BR, et al. 2007. J. Immunol. 178:5366. (FC)
13. Patole PS, et al. 2005. J. Am. Soc. Nephrol. 16:3273. PubMed
14. Wu S, et al. 2005. Neurosci Lett. 394:158. PubMed
15. Carlson MJ, et al. 2009. Blood 113:1365. PubMed

PMA+ Ionomycin-stimulated (6 hours) C57BL/6

PMA+ Ionomycin-stimulated (6 hours) C57BL/6 mouse splenocytes (in the presence of monensin) were surface stained with CD3 FITC and then intracellularly stained with TNF-α (clone MP6-XT22) Brilliant Violet 650™.



Description:

TNF-α is secreted by macrophages, monocytes, neutrophils, T-cells (principally CD4+), and NK-cells. Many transformed cell lines also secrete TNF-α. Monomeric mouse TNF-α is a 156 amino acid protein (N-glycosylated) with a reported molecular weight of 17.5 kD. TNF-α forms multimeric complexes; stable trimers are most common in solution. A 26 kD membrane form of TNF-α has also been described. TNF-α binding to surface receptors elicits a wide array of biologic activities including: cytolysis and cytostasis of many tumor cell lines in vitro, hemorrhagic necrosis of tumors in vivo, increased fibroblast proliferation, and enhanced chemotaxis and phagocytosis in neutrophils.

Other Names: Tumor necrosis factor-α, Cachectin, Necrosin, Macrophage cytotoxic factor (MCF), Differentiation inducing factor (DIF), TNFSF-2, TNF-a, TNF-alpha
Structure: TNF superfamily; dimer/trimer; 17.5-150 kD (Mammalian)
Regulation: Processed by TACE for secretion; upregulated by interferons, IL-2, GM-CSF, substance P, bradykinin, PAF, immune complexes, and cyclooxygenase; downregulated by IL-6, TGF-β, vitamin D3, prostaglandin E2, and PAF antagonists
Cellular Sources: Activated monocytes, neutrophils, macrophages, T cells, B cells, NK cells, LAK cells
Cellular Targets: Monocytes, neutrophils, macrophages, T cells, fibroblasts, endothelial cells, osteoclasts, adipocytes, astroglia, microglia
Receptors: TNFRSF1A (TNF-R1, CD120a, TNFR-p60 Type β, p55); TNFRSF1B (TNF-R2, CD120b, TNFR-p80 Type A, p75)
Bioactivity/Activities: Paracrine/endocrine mediator of inflammatory and immune functions; selectively cytotoxic for transformed cells; endothelial cell alterations; chemoattractant
Antigen References:

1. Fitzgerald K, et al. Eds. 2001. The Cytokine FactsBook. Academic Press, San Diego.
2. Beutler B, et al. 1988. Annu. Rev. Biochem. 57:505.
3. Beutler B, et al. 1989. Annu. Rev. Immunol. 7:625.
4. Tracey K, et al. 1993. Crit. Care Med. 21:S415.