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Ultra-LEAF™ Purified 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
PMA/Ionomycin-stimulated BALB/c T cells were stained with MP6-XT22 PE
  • mp6-xt22
    PMA/Ionomycin-stimulated BALB/c T cells were stained with MP6-XT22 PE
  • MP6-XT22_013111
    Immortalized murine bone marrow-derived macrophages stimulated overnight with LPS were stained with Atto-488 phalloidin (green) and purified TNF-α (clone MP6-XT22), secondarily stained with Goat anti-Rat IgG Dylight 594 (red). Data provided by James Harris, Trinity College.
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506331 100 µg 146€
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506332 1 mg 507€
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506347 5 mg 595€
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506348 25 mg 1760€
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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
0.2 µm filtered in phosphate-buffered solution, pH 7.2, containing no preservative.
Endotoxin Level
Less than 0.01 EU/µg of the protein (< 0.001 ng/µg of the protein) as determined by the LAL test.
Preparation
The Ultra-LEAF™ (Low Endotoxin, Azide-Free) antibody was purified by affinity chromatography.
Concentration
The antibody is bottled at the concentration indicated on the vial, typically between 2 mg/mL and 3 mg/mL. Older lots may have also been bottled at 1 mg/mL. To obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C. This Ultra-LEAF™ solution contains no preservative; handle under aseptic conditions.
Application

ELISA, ICFC - Quality tested
CyTOF®, ICC - Verified
IHC-F, WB, Neut - Reported in the literature, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by ELISA assay. For ELISA capture applications, a concentration range of 2.0 - 6.0 µg/mL is recommended. To obtain a linear standard curve, serial dilutions of mouse TNF-α recombinant protein ranging from 500 to 4 pg/mL are recommended for each ELISA plate. It is recommended that the reagent be titrated for optimal performance for each application.

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
  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. Qi S, et al. 2019. J Immunol. 203:3045. PubMed
  2. McKnight Q, et al. 2020. J Bone Miner Res. 35:1352. PubMed
  3. Mabire M, et al. 2023. Nat Commun. 14:1830. PubMed
  4. Peng L, et al. 2024. Macromol Rapid Commun. e2300559:45. PubMed
  5. Carlson M, et al. 2017. Blood. 113(6):1365-74. PubMed
  6. Kearney C, et al. 2017. Cell Death Differ.. 10.1038/cdd.2017.94. PubMed
  7. Pham THM, et al. 2020. Cell Host & Microbe. 27(1):54-67.e5.. PubMed
  8. Wood M, Rios D, Williams I 2016. Am J Physiol Cell Physiol. 311: C498 - C507. PubMed
  9. Nie X et al. 2018. Neuron. 99(3):464-479 . PubMed
  10. Iizasa E, et al. 2021. Nat Commun. 12:2299. PubMed
  11. Jung H, et al. 2019. Autophagy. 15:1990. PubMed
  12. Kurashima Y, et al. 2021. Nat Commun. 12:1067. PubMed
  13. Jain A, et al. 2020. Nat Immunol. 0.920138889. PubMed
RRID
AB_2616671 (BioLegend Cat. No. 506331)
AB_2616671 (BioLegend Cat. No. 506332)
AB_2616671 (BioLegend Cat. No. 506347)
AB_2616671 (BioLegend Cat. No. 506348)

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

Do you guarantee that your antibodies are totally pathogen free?

BioLegend does not test for pathogens in-house aside from the GoInVivo™ product line. However, upon request, this can be tested on a custom basis with an outside, independent laboratory.

Does BioLegend test each Ultra-LEAF™ antibody by functional assay?

No, BioLegend does not test Ultra-LEAF™ antibodies by functional assays unless otherwise indicated. Due to the possible complexities and variations of uses of biofunctional antibodies in different assays and because of the large product portfolio, BioLegend does not currently perform functional assays as a routine QC for the antibodies. However, we do provide references in which the antibodies were used for functional assays and we do perform QC to verify the specificity and quality of the antibody based on our strict specification criteria.

Does BioLegend test each Ultra-LEAF™ antibody for potential pathogens?

No, BioLegend does not test for pathogens in-house unless otherwise indicated.  However, we can recommend an outside vendor to perform this testing as needed.

Have you tested this Ultra-LEAF™ antibody for in vivo or in vitro applications?

We don't test our antibodies for in vivo or in vitro applications unless otherwise indicated. Depending on the product, the TDS may describe literature supporting usage of a particular product for bioassay. It may be best to further consult the literature to find clone specific information.

Go To Top Version: 3    Revision Date: 01-20-2021

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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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

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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