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
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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506301 50 µg $83.00
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506302 500 µg $235.00
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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.

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

ELISA, ICFC - Quality tested
CyTOF®, ICC - Verified
IHC-F, WB - 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

(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. Wolf Y, et al. 2017. J Exp Med. 214:905. PubMed
  2. Yan J, et al. 2020. Cancer Discov. 10:124. PubMed
  3. Yu H, et al. 2020. Front Immunol. 11:1773. PubMed
  4. Kim M, et al. 2021. Exp Mol Med. 53:67. PubMed
  5. Bahreyni A, et al. 2023. Biomed Pharmacother. 163:114789. PubMed
  6. Bahreyni A, et al. 2023. BMC Med. 21:193. PubMed
  7. Asthagiri Arunkumar G, et al. 2019. Vaccine. 37:5567. PubMed
  8. Chartrand K, et al. 2018. Front Immunol. 1.642361111. PubMed
  9. Wang W, et al. 2018. Cancer Cell. 34:757. PubMed
  10. Kapadia D, et al. 2011. PLoS One. 6:e19376. PubMed
  11. Wu S, et al. 2006. Neurosci Lett. 394:158. PubMed
  12. Sakai J, et al. 2017. Sci Rep. 1.283333333. PubMed
  13. Knuschke T, et al. 2018. Front Immunol. 0.801388889. PubMed
  14. Mouat IC, et al. 2021. Elife. 10:. PubMed
  15. Patole P, et al. 2005. J Am Soc Nephrol. 2.939583333. PubMed
  16. Al Sayed MF, et al. 2019. Cancer Res. 79:346. PubMed
  17. Tan Z, et al. 2020. Mol Ther Oncolytics. 16:302. PubMed
  18. Hu G, et al. 2021. Nat Commun. 12:773. PubMed
  19. Enders M, et al. 2020. J Immunol. 204:87. PubMed
  20. Borriello F, et al. 2018. Front Immunol. 8:1772. PubMed
  21. Dineen S, et al. 2010. Cancer Res . 70:2852. PubMed
  22. Johnson SA, et al. 2021. Eur J Immunol. 51:3228. PubMed
  23. Engdahl C, et al. 2018. Arthritis Res Ther. 20:84. PubMed
  24. Daley D, et al. 2017. Nat Med. 23:556. PubMed
  25. Shin H, et al. 2017. PLoS One.. 10.1371/journal.ppat.1006544. PubMed
  26. Muhammad F, et al. 2020. J Autoimmun. 111:102441. PubMed
  27. Ranjan K, et al. 2021. J Clin Invest. 131:. PubMed
  28. Kakaradov B, et al. 2017. Nat Immunol. 18:422. PubMed
  29. Scortegagna M, et al. 2020. Nat Commun. 11:99. PubMed
  30. Uehara H, et al. 2021. eLife. 10:00. PubMed
  31. Stelekati E, et al. 2018. Cell Rep. 2.445833333. PubMed
  32. Diao J, et al. 2018. J Immunol. 201:1306. PubMed
  33. Pushalkar S, et al. 2018. Cancer Discov. 0.613194444. PubMed
  34. Naito H, et al. 2019. Dev Cell. 48:151. PubMed
RRID
AB_315423 (BioLegend Cat. No. 506301)
AB_315423 (BioLegend Cat. No. 506302)

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: 4    Revision Date: 01/20/2021

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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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