Key Product Details

Species Reactivity

Human, Rat, Mouse (Negative)

Applications

Western Blot, Flow Cytometry, Immunofluorescence, Immunocytochemistry/ Immunofluorescence, CyTOF-ready

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # J2D10

Format

Azide and BSA Free
View all TNF-alpha Primary Antibodies »

Product Specifications

Immunogen

Recombinant human TNF-alpha (Uniprot: P01375)

Reactivity Notes

Does not react with Mouse.

Localization

Cytoplasmic and extracellular (secreted)

Specificity

This antibody neutralizes Hu rTNFa mediated cytotoxicity of L929 cells and inhibits tumor growth in mice. It protects mice against toxicity of HurTNFa. Tumor Necrosis Factor Alpha (TNF-alpha) is a protein secreted by lipopolysaccharide-stimulated macrophages, and causes tumor necrosis when injected into tumor bearing mice. TNF-alpha is believed to mediate pathogenic shock and tissue injury associated with endotoxemia. TNF-alpha exists as a multimer of two, three, or five non-covalently linked units, but shows a single 17kDa band following SDS PAGE under non-reducing conditions. TNF-alpha is closely related to the 25kDa protein Tumor Necrosis Factor beta (lymphotoxin), sharing the same receptors and cellular actions. TNF-alpha causes cytolysis of certain transformed cells, being synergistic with interferon gamma in its cytotoxicity. Although it has little effect on many cultured normal human cells, TNF-alpha appears to be directly toxic to vascular endothelial cells. Other actions of TNF-alpha include stimulating growth of human fibroblasts and other cell lines, activating polymorphonuclear neutrophils and osteoclasts, and induction of interleukin 1, prostaglandin E2 and collagenase production.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Theoretical MW

17 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Description

1.0 mg/ml of antibody purified by Protein L. Prepared in 10mM PBS WITHOUT BSA. Also available at 200 ug/ml WITH BSA & azide (NBP2-45333).

Antibody with azide - store at 2 to 8C. Antibody without azide - store at -20 to -80C.

Scientific Data Images for TNF-alpha Antibody (J2D10) - Azide and BSA Free

TNF-alpha Antibody (J2D10) - Azide and BSA Free Western Blot: TNF-alpha Antibody (J2D10) - Azide and BSA Free [NBP2-47677] -

Western Blot: TNF-alpha Antibody (J2D10) - Azide and BSA Free [NBP2-47677] -

Western Blot: TNF-alpha Antibody (J2D10) - Azide and BSA Free [NBP2-47677] - Analysis of pro-inflammatory signalling in left ventricle lysates. A-F, quantification and G, representative immunoblots and total protein staining of procsp1, csp1, proIL-1 beta, IL-1 beta and TNF in sham-operated group (Sham), non-infarcted (HFni) and infarcted (HFi) tissue of failing hearts. Data are presented as mean ± SEM; n = 6-9 per group; *P < 0.05 vs Sham; #P < 0.05 vs HFni
TNF-alpha Antibody (J2D10) - Azide and BSA Free Western Blot: TNF-alpha Antibody (J2D10) - Azide and BSA Free [NBP2-47677] -

Western Blot: TNF-alpha Antibody (J2D10) - Azide and BSA Free [NBP2-47677] -

Western Blot: TNF-alpha Antibody (J2D10) - Azide and BSA Free [NBP2-47677] - Analysis of pro-inflammatory signalling in left ventricle lysates. Representative immunoblots and total protein staining of procsp1, csp1, proIL-1beta, IL-1beta and TNF-alpha in sham-operated group (Sham), non-infarcted (HFni) and infarcted (HFi) tissue of failing hearts. Data are presented as mean +/- SEM; n = 6-9 per group; *P < 0.05 vs Sham; #P < 0.05 vs HFni Image collected and cropped by CiteAb from the following publication (https://onlinelibrary.wiley.com/doi/abs/10.1111/jcmm.14536), licensed under a CC-BY license.

Applications for TNF-alpha Antibody (J2D10) - Azide and BSA Free

Application
Recommended Usage

Flow Cytometry

0.5 - 1 ug/million cells in 0.1 ml

Immunocytochemistry/ Immunofluorescence

1-2 ug/ml

Immunofluorescence

1 - 2 ug/ml
Application Notes
Inhibits Tumor Growth: Order Ab without BSA/Azide.
Neutralizes TNF-alpha- Mediated Cytotoxicity: Order Ab without BSA/Azide.
Protects Mice Against Toxicity of TNF-alpha. Order Ab without BSA/Azide.
Optimal dilution for a specific application should be determined.

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein L purified

Formulation

10 mM PBS

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20 to -80C. Avoid freeze-thaw cycles.

Background: TNF-alpha

Tumor necrosis factor (TNF)-alpha is a pro-inflammatory cytokine belonging to the TNF superfamily that is secreted by monocytes/macrophages, T cell, and natural killer (NK), among others (1). TNF-alpha is synthesized as a 233 amino acid (aa) transmembrane protein (mTNF-alpha) with a theoretical molecular weight (MW) of 26 kDa (1,2) that forms a homotrimer. mTNF-alpha is cleaved by TNF-alpha converting enzyme (TACE) and released in its 157 aa, 17 kDa soluble form (sTNF-alpha) (1-5). Both mTNF-alpha and sTNF-alpha are capable of binding type 1 TNF receptors (TNFR1), whereas mTNF-alpha predominately binds to TNFR2 (1,2). TNF-alpha binding to its receptors causes receptor recruitment of adaptor proteins, formation of signaling complexes, and downstream signaling cascades (e.g. MAPK, NF-kappaB, and Caspase-8), leading to distinct cellular responses such as survival, proliferation, inflammation, necroptosis, and apoptosis (1-5).

TNF-alpha is critical for normal immune response; however, dysregulation of TNF-alpha production can result in various pathologies (2,4,5). Excessive production of pro-inflammatory cytokines including interleukin 1 (IL-1), IL-6, and TNF-alpha has been implicated in an array of autoimmune diseases like rheumatoid arthritis (RA), inflammatory bowel disease (IBD), and psoriasis (2,4,5). Anti-TNF monoclonal antibodies, including Infliximab, and soluble TNFR have been approved for the treatment of autoimmune and TNF-mediated diseases (5). Additionally, data suggests that TNF inhibitors can be beneficial for treating patients experiencing immune-related adverse events associated with immune checkpoint inhibitor cancer treatment (6).

References

1. Holbrook J, Lara-Reyna S, Jarosz-Griffiths H, McDermott M. Tumour necrosis factor signalling in health and disease. F1000Res. 2019;8:F1000 Faculty Rev-111. https://doi.org/10.12688/f1000research.17023.1

2. Jang DI, Lee AH, Shin HY, et al. The Role of Tumor Necrosis Factor Alpha (TNF-alpha) in Autoimmune Disease and Current TNF-alpha Inhibitors in Therapeutics. Int J Mol Sci. 2021;22(5):2719. https://doi.org/10.3390/ijms22052719

3. Horiuchi T, Mitoma H, Harashima S, Tsukamoto H, Shimoda T. Transmembrane TNF-alpha: structure, function and interaction with anti-TNF agents. Rheumatology (Oxford). 2010;49(7):1215-1228. https://doi.org/10.1093/rheumatology/keq031

4. Webster JD, Vucic D. The Balance of TNF Mediated Pathways Regulates Inflammatory Cell Death Signaling in Healthy and Diseased Tissues. Front Cell Dev Biol. 2020;8:365. https://doi.org/10.3389/fcell.2020.00365

5. Kalliolias GD, Ivashkiv LB. TNF biology, pathogenic mechanisms and emerging therapeutic strategies. Nat Rev Rheumatol. 2016; 12(1):49-62. https://doi.org/10.1038/nrrheum.2015.169

6. Chen AY, Wolchok JD, Bass AR. TNF in the era of immune checkpoint inhibitors: friend or foe?. Nat Rev Rheumatol. 2021;17(4):213-223. doi:10.1038/s41584-021-00584-4

Long Name

Tumor Necrosis Factor alpha

Alternate Names

Cachetin, DIF, TNF, TNF-A, TNFA, TNFalpha, TNFG1F, TNFSF1A, TNFSF2

Gene Symbol

TNF

Additional TNF-alpha Products

Product Documents for TNF-alpha Antibody (J2D10) - Azide and BSA Free

Certificate of Analysis

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Product Specific Notices for TNF-alpha Antibody (J2D10) - Azide and BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for TNF-alpha Antibody (J2D10) - Azide and BSA Free

Showing  1 - 4 of 4 FAQs Showing All

    • Q: Can your TNF-alpha products be used to treat TBI victims and therefore avoid the perispinal injections with Enbrel?
      A: I am very sorry, but all of our products are for scientific research use only, and none are intended or approved for use in humans.

    • Q: I am interested in a TNF alpha antibody, cross reactive for human, rat and mouse (host species: rabbit). Could your product NBP1-19532 be used in western blotting applications? Or do you have a similar product in your catalog which could fit with my request?
      A: The antibody you mention, NBP1-19532, has not yet been validated in Western blot. I would instead recommend either NBP1-67821 or NB600-587. These are both rabbit polyclonal antibodies that cross-reacts with human, mouse and rat and have been used in Western blotting.

    • A: We currently have two TNFalpha ELISA kits specific for mouse: catalog numbers KA0257 and NBP1-92670.

    • Q: I would like to ask you for help. I need an antibody for Elisa to detect human TNF alpha in a supernatant from a cell culture (meaning supernatant after centrifugation of collected cell suspension from a plate well). Which of your antibodies against human tnf alpha would be suitable? I would like to buy only the primary antibody.
      A: I would recommend catalog number NBP1-67821 or NB600-587; however, a full list of our anti-human TNF alpha antibodies suitable for use in ELISA can be found using this link.
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