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Safety Data Sheet (SDS) Pacific Blue Anti-mouse TNF-alpha Antibody     Product Data Sheet (PDF)    
Pacific Blue™ Anti-mouse TNF-α Antibody
3131590 100 µg $220.00       
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.
Preparation: The antibody was purified by affinity chromatography, and conjugated with Pacific Blue™ under optimal conditions. The solution is free of unconjugated Pacific Blue™.
Concentration: 0.5 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 ≤ 0.25 µg per 106 cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.

* Pacific Blue™ has a maximum emission of 455 nm when it is excited at 405 nm. Prior to using Pacific Blue™ conjugate for flow cytometric analysis, please verify your flow cytometer's capability of exciting and detecting the fluorochrome.

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 C57BL/6 mouse

PMA + Ionomycin-stimulated C57BL/6 mouse splenocytes (in the presence of monensin) were stained with CD3 PE, fixed, permeabilized and then stained with TNF-α (clone MP6-XT22) Pacific Blue™ (top) or rat IgG1, κ Pacific Blue™ isotype control (bottom).





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.


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