TNF-alpha Antibody (J2D10) [Alexa Fluor® 350]

TNF-alpha Antibody (J2D10) [Alexa Fluor® 350] [NBP2-47677AF350] - Vial of Alexa Fluor 350 conjugated antibody. Alexa Fluor 350 is optimally excited at 346 nm by the UV laser (350 or 355 nm) and has an emission maximum of 442 nm.

• Reactivity: Hu, Rt, Mu(-)
• Applications: WB, Flow, ICC/IF, CyTOF-ready, ICC/IF
• Clone: J2D10
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Alexa Fluor 350

TNF-alpha Antibody (J2D10) [Alexa Fluor® 350] Summary

• Immunogen: Recombinant human TNF-alpha (Uniprot: P01375)
• 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.
• Isotype: IgG1 Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: TNF
• Purity: Protein A or G purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Flow Cytometry
  • Immunocytochemistry/ Immunofluorescence
  • Immunofluorescence
  • Western Blot
• Application Notes: Optimal dilution of this antibody should be experimentally determined.

Reactivity Notes

Does not react with Mouse.

Packaging, Storage & Formulations

• Storage: Store at 4C in the dark.
• Buffer: 50mM Sodium Borate
• Preservative: 0.05% Sodium Azide
• Purity: Protein A or G purified

Notes

Alexa Fluor (R) products are provided under an intellectual property license from Life Technologies Corporation. The purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: (i) in manufacturing; (ii) to provide a service, information, or data in return for payment; (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com. This conjugate is made on demand. Actual recovery may vary from the stated volume of this product. The volume will be greater than or equal to the unit size stated on the datasheet.

Alternate Names for TNF-alpha Antibody (J2D10) [Alexa Fluor® 350]

• APC1 protein
• Cachectin
• Cachetin
• DIF
• TNF
• TNF, monocyte-derived
• TNFA
• TNF-A
• TNFalpha
• TNF-alpha
• TNF-alphacachectin
• TNFATNF, macrophage-derived
• TNFG1F
• TNFSF1A
• TNFSF2
• TNFSF2TNF superfamily, member 2
• tumor necrosis factor (TNF superfamily, member 2)
• tumor necrosis factor alpha
• Tumor necrosis factor ligand superfamily member 2
• tumor necrosis factor
• tumor necrosis factor-alpha

Background

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

Limitations

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.

FAQs for TNF-alpha Antibody (NBP2-47677AF350). (Showing 1 - 4 of 4 FAQ).

1. I need to know about TNF-alpha antibody kits for mice.
   • We currently have two TNFalpha ELISA kits specific for mouse: catalog numbers KA0257 and NBP1-92670.
2. 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.
   • 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.
3. 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?
   • 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.
4. Can your TNF-alpha products be used to treat TBI victims and therefore avoid the perispinal injections with Enbrel?
   • 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.

Bioinformatics

• Gene Symbol: TNF