- MP6-XT22 (See other available formats)
- Other Names
- Tumor necrosis factor-α, Cachectin, Necrosin, Macrophage cytotoxic factor (MCF), Differentiation inducing factor (DIF), TNFSF-2, TNF-a, TNF-alpha
- Rat IgG1, κ
- Ave. Rating
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- Product Citations
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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.Product Details
- Antibody Type
- Host Species
- E. coli-expressed, recombinant mouse TNF-α
- 0.2 µm filtered in phosphate-buffered solution, pH 7.2, containing no preservative. Endotoxin level is <0.1 EU/µg of the protein (<0.01 ng/µg of the protein) as determined by the LAL test.
- The LEAF™ (Low Endotoxin, Azide-Free) antibody was purified by affinity chromatography.
- 1.0 mg/ml
- Storage & Handling
- The TNF-α antibody solution should be stored undiluted between 2°C and 8°C. This LEAF™ solution contains no preservative; handle under aseptic conditions.
ELISA, ICFC - Quality tested
CyTOF®, IF - Validated
IHC, WB, Neut - Reported in the literature
- Recommended Usage
Each lot of this antibody is quality control tested by ELISA assay. For ELISA capture applications, a concentration range of 2-6 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. No. 510802/510804) 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.
Neutralization1,5,10,16,17: 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 to 430906) are specially developed and recommended.
(PubMed link indicates BioLegend citation)
- Abrams J, et al. 1992. Immunol. Rev. 127:5. (Neut)
- Abrams J, et al. 1995. Curr. Prot. Immunol. John Wiley and Sons, New York. Unit 6.20
- Mo X, et al. 1995. J. Virol. 69:1288.
- Sarawar S, et al. 1994. J. Immunol. 153:1246.
- Via C, et al. 2001. J. Immunol. 167:6821. (Neut)
- Infante-Duarte C, et al. 2000 J. Immunol. 165:6107. (FC)
- Jacobs M, et al. 2000. Immunology 100:494. (IHC)
- Marinova-Mutachieva L, et al. 1997. Clin. Exp. Immunol. 107:507. (IHC)
- Williams RO, et al. 2000. J. Immunol. 165:7240. (IHC)
- Scanga CA, et al. 1999. Infect. Immun. 67:4531. (Neut)
- Akilov OE, et al. 2007. J. Leukoc. Biol. 2007;10.1189/jlb.0706439. (FC)
- Lawson BR, et al. 2007. J. Immunol. 178:5366. (FC)
- Patole PS, et al. 2005. J. Am. Soc. Nephrol. 16:3273. PubMed
- Wu S, et al. 2005. Neurosci Lett. 394:158. PubMed
- Carlson MJ, et al. 2009. Blood 113:1365. PubMed
- Shivakumar P, et al. 2017. JCI Insight. 2:e88747 1. PubMed
- Kearney CJ, et al. 2017. Cell Death Differ. 10.1038/cdd.2017.94. PubMed
- Product Citations
AB_315430 (BioLegend Cat. No. 506309)
AB_315431 (BioLegend Cat. No. 506310)
AB_2204355 (BioLegend Cat. No. 506325)
- TNF superfamily; dimer/trimer; 17.5-150 kD (Mammalian)
- 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
- TNFRSF1A (TNF-R1, CD120a, TNFR-p60 Type β, p55); TNFRSF1B (TNF-R2, CD120b, TNFR-p80 Type A, p75)
- Cell Type
- Biology Area
- Immunology, Innate Immunity
- Molecular Family
- 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.
- 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
- View information about TNF-alpha on UniProt.org
- Does BioLegend test each LEAF™ antibody by functional assay?
No, BioLegend does not test 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.
- 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 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 LEAF™/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.
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