Purified anti-mouse TNF-α (Maxpar® Ready) 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_Purified_TNFa_CyTOF_1_112113
C57BL/6 mouse splenocytes were incubated for 6 hours in media alone (top) or with PMA and Ionomycin (bottom) in the presence of monensin and brefeldin A. Cells were then fixed, permeabilized, and stained with 162Dy-anit-TNFα (MP6-XT22). CD4+ T lymocyte are displayed in the analysis. Data provided by DVS Sciences.
  • MP6-XT22_Purified_TNFa_CyTOF_1_112113
    C57BL/6 mouse splenocytes were incubated for 6 hours in media alone (top) or with PMA and Ionomycin (bottom) in the presence of monensin and brefeldin A. Cells were then fixed, permeabilized, and stained with 162Dy-anit-TNFα (MP6-XT22). CD4+ T lymocyte are displayed in the analysis. Data provided by DVS Sciences.
  • MP6-XT22_Purified_TNFa_CyTOF_2_112113
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506335 100 µg 141 CHF
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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 (pdf)

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 EDTA.
Preparation
The antibody was purified by affinity chromatography.
Concentration
1.0 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

ELISA - Quality tested
CyTOF® - Verified

Recommended Usage

This product is suitable for use with the Maxpar® Metal Labeling Kits. For metal labeling using Maxpar® Ready antibodies, proceed directly to the step to Partially Reduce the Antibody by adding 100 µl of Maxpar® Ready antibody to 100 µl of 4 mM TCEP-R in a 50 kDa filter and continue with the protocol. Always refer to the latest version of Maxpar® User Guide when conjugating Maxpar® Ready antibodies.

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.

Additional Product Notes

Maxpar® is a registered trademark of Standard BioTools Inc.

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. McDonald B, et al. 2020. Cell Host Microbe. 28(5):660-668.e4. PubMed
RRID
AB_2562848 (BioLegend Cat. No. 506335)

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

Related FAQs

Can I obtain CyTOF data related to your Maxpar® Ready antibody clones?

We do not test our antibodies by mass cytometry or on a CyTOF machine in-house. The data displayed on our website is provided by Fluidigm®. Please contact Fluidigm® directly for additional data and further details.

http://techsupport.fluidigm.com/

Can I use Maxpar® Ready format clones for flow cytometry staining?

We have not tested the Maxpar® Ready antibodies formulated in solution containing EDTA for flow cytometry staining. While it is likely that this will work in majority of the situations, it is best to use the non-EDTA formulated version of the same clone for flow cytometry testing. The presence of EDTA in some situations might negatively affect staining.

I am having difficulty observing a signal after conjugating a metal tag to your Maxpar® antibody. Please help troubleshoot.

We only supply the antibody and not test that in house. Please contact Fluidigm® directly for troubleshooting advice: http://techsupport.fluidigm.com/

Is there a difference between buffer formulations related to Maxpar® Ready and purified format antibodies?

The Maxpar® Ready format antibody clones are formulated in Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and EDTA. The regular purified format clones are formulated in solution that does not contain any EDTA. Both formulations are however without any extra carrier proteins.

Go To Top Version: 3    Revision Date: 07.22.2022

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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