- 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
- Submit a Review
- Product Citations
- publications
![MP6-XT22 MP6-XT22](https://d1spbj2x7qk4bg.cloudfront.net/Admin/Public/GetImage.ashx?Image=/Files/Images/media_assets/products/product_images/mp6-xt22.jpg&Width=240&Height=300&altFmImage_path=&Compression=90&Crop=5)
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PMA/Ionomycin-stimulated BALB/c T cells were stained with MP6-XT22 PE -
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.
Cat # | Size | Price | Quantity Check Availability | Save | ||
---|---|---|---|---|---|---|
506301 | 50 µg | $83 | ||||
506302 | 500 µg | $235 |
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 DetailsProduct Details
- 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
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ELISA, ICFC - Quality tested
CyTOF®, ICC - Verified
IHC-F, WB - Reported in the literature, not verified in house - Recommended Usage
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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) -
- 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
- RRID
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AB_315422 (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
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Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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