The antibody was purified by affinity chromatography, and conjugated with Pacific Blue™ under optimal conditions. The solution is free of unconjugated Pacific Blue™.
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.
ICFC - Quality tested
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.
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.
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 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).
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.
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
Activated monocytes, neutrophils, macrophages, T cells, B cells, NK cells, LAK cells
TNFRSF1A (TNF-R1, CD120a, TNFR-p60 Type β, p55); TNFRSF1B (TNF-R2, CD120b, TNFR-p80 Type A, p75)
Paracrine/endocrine mediator of inflammatory and immune functions; selectively cytotoxic for transformed cells; endothelial cell alterations; chemoattractant
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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