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Brilliant Violet 650™ anti-mouse TNF-α Antibody

Pricing & Availability
Clone
MP6-XT22 (See other available formats)
Regulatory Status
RUO
Other Names
Tumor necrosis factor-α, Cachectin, Necrosin, Macrophage cytotoxic factor (MCF), Differentiation inducing factor (DIF), TNFSF-2, TNF-a, TNF-alpha
Isotype
Rat IgG1, κ
Ave. Rating
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Product Citations
publications
MP6-XT22_BV650_TNF-a_Antibody_FC_082913.jpg
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™.
  • MP6-XT22_BV650_TNF-a_Antibody_FC_082913.jpg
    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™.
Compare all formats See Brilliant Violet 650™ spectral data
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506333 50 µg $273
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Description

TNF-α is secreted by macrophages, monocytes, neutrophils, T-cells, 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.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Mouse
Antibody Type
Monoclonal
Host Species
Rat
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.
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
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.


Learn more about Brilliant Violet™.

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.
Excitation Laser
Violet Laser (405 nm)
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 (Cat. Nos. 510802 & 510804) as the capture antibody.
ELISA Capture: The purified MP6-XT22 antibody is useful as the capture antibody in a sandwich ELISA when used in conjunction with the biotinylated Poly5160 antibody (Cat. No. 516003) as the detection antibody and recombinant mouse TNF-α (Cat. No. 575209) as the standard.
Flow Cytometry6,11,12:The fluorochrome-labeled MP6-XT22 antibody is useful for intracellular immunofluorescent staining and flow cytometric analysis to identify TNF-a-producing cells within mixed cell populations.
Neutralization1,5,10,16,17:The MP6-XT22 antibody can neutralize the bioactivity of natural or recombinant TNF-α. The LEAF™ purified antibody (Endotoxin < 0.1 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for neutralization of mouse TNF-α bioactivity in vivo and in vitro(Cat. No. 506310). For in vivo studies or highly sensitive assays, we recommend Ultra-LEAF™ purified antibody (Cat. No. 506332) with a lower endotoxin limit than standard LEAF™ purified antibodies (Endotoxin < 0.01 EU/µg).
Additional reported applications (for the relevant formats) include: Western blotting, immunohistochemical staining of paraformaldehyde-fixed, saponin-treated frozen tissue sections7-9 in vivo detection5, immunofluorescence, and immunocytochemistry.
Note: For testing mouse TNF-α in serum, plasma or supernatant, BioLegend's ELISA Max™ Sets (Cat. No. 430901) are specially developed and recommended.

Application References

(PubMed link indicates BioLegend citation)
  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
  16. Shivakumar P, et al. 2017. JCI Insight. 2:e88747 1. PubMed
  17. Kearney CJ, et al. 2017. Cell Death Differ. 10.1038/cdd.2017.94. PubMed
Product Citations
  1. Shakya S, et al. 2020. MethodsX. 7:101027. PubMed
  2. Marchelletta RR, et al. 2021. J Clin Invest. 131:. PubMed
  3. D'Angeli V, et al. 2022. Eur J Immunol. 52:1058. PubMed
  4. Lin YH 2023. Immunity. 56(1):207-223.e8. PubMed
  5. Matter AL, et al. 2022. J Immunol. 209:1880. PubMed
  6. Rodriguez-García A, et al. 2021. Nat Commun. 12:877. PubMed
  7. Sharma NS, et al. 2020. J Clin Invest. 130:451. PubMed
  8. Hering L, et al. 2021. Int J Mol Sci. 22:. PubMed
  9. Palathingal Bava E, et al. 2022. JCI Insight. 7:. PubMed
  10. Shi L, et al. 2016. Nat Commun. 7:12335. PubMed
  11. Rosenbaum SR, et al. 2020. Cell Rep. 30:510. PubMed
  12. Breznik JA, et al. 2021. Immunohorizons. 5:370. PubMed
  13. Jablonski K, et al. 2015. PLoS One. 10: 0145342. PubMed
  14. Kung YJ, et al. 2022. J Biomed Sci. 29:57. PubMed
  15. Hering L, et al. 2020. Front Immunol. 1.747222222. PubMed
  16. Kumar R, et al. 2020. Cell Reports. 30(8):2512-2525. PubMed
  17. Fearon AE, et al. 2021. iScience. 24:103143. PubMed
RRID
AB_2562450 (BioLegend Cat. No. 506333)

Antigen Details

Structure
TNF superfamily; dimer/trimer; 17.5-150 kD (Mammalian)
Bioactivity
Paracrine/endocrine mediator of inflammatory and immune functions; selectively cytotoxic for transformed cells; endothelial cell alterations; chemoattractant
Cell Sources
Activated monocytes, neutrophils, macrophages, T cells, B cells, NK cells, LAK cells
Cell 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)
Cell Type
Tregs
Biology Area
Immunology, Innate Immunity
Molecular Family
Cytokines/Chemokines
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.

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
Gene ID
21926 View all products for this Gene ID
UniProt
View information about TNF-alpha on UniProt.org
Go To Top Version: 1    Revision Date: 09/03/2013

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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