TNF-alpha Antibody (28401) [Unconjugated]

Western blot shows 25 ng of Recombinant Human TNF‑ alpha (Catalog # 210-TA), Recombinant Mouse TNF‑ alpha aa 80-235 (Catalog # 410-MT), Recombinant Rat TNF‑ alpha (Catalog # 510-RT), and Recombinant Human APRIL/TNFSF13 (Catalog # 5860-AP). PVDF Membrane was probed with 1 µg/mL of Mouse Anti-Human TNF‑ alpha Monoclonal Antibody (Catalog # MAB610) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for TNF‑ alpha at approximately 15 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 3.

Recombinant Human TNF‑ alpha (Catalog #210‑TA) induces cytotoxicity in the the L‑929 mouse fibroblast cell line in a dose-dependent manner (orange line). Cytotoxicity elicited by Recombinant Human TNF‑ alpha (0.25 ng/mL) is neutral­ized (green line) by increasing concentrations of Mouse Anti-Human TNF‑ alpha Monoclonal Antibody (Catalog # MAB610). The ND50 is typically0.01‑0.04 µg/mL in the presence of the metabolic inhibitor actinomycin D.

TNF‑ alpha was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) stimulated with LPS and monensin using Mouse Anti-Human TNF‑ alpha Monoclonal Antibody (Catalog # MAB610) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (yellow; Catalog # NL007) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

• Reactivity: Hu
• Applications: WB, B/N, ELISA(Cap), ICC/IF
• Clone: 28401
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

TNF-alpha Antibody (28401) [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human TNF-alpha
  Gly57-Leu233 (predicted)
  Accession # P01375
• Specificity: Detects human TNF-alpha in ELISAs and Western blots. In sandwich ELISAs, less than 0.05% cross-reactivity with recombinant human TNF‑ beta, recombinant mouse TNF‑ alpha, recombinant rat TNF‑ alpha, and recombinant porcine TNF‑ alpha is observed.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: TNF
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Western Blot 1 ug/mL
  • ELISA Capture (Matched Antibody Pair) 2-8 ug/mL
  • ELISA Detection (Matched Antibody Pair) 0.1-0.4 ug/mL
  • Immunocytochemistry 8-25 ug/mL
  • Neutralization 0.01-0.04 ug/mL
• Application Notes: ELISA Detection: Human TNF-alpha Biotinylated Antibody (Catalog number BAF210)
  Standard: Recombinant Human TNF-alpha (Catalog number 210-TA)

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for TNF-alpha Antibody (28401) [Unconjugated]

• 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 alpha (TNF-alpha, TNF- alpha, TNFA ), also known as Cachectin and TNFSF2, is the prototypic ligand of the TNF superfamily. It is a pleiotropic molecule that plays a central role in inflammation, immune system development, apoptosis, and lipid metabolism. TNF- is produced by several lymphoid cells as well as by astrocytes, endothelial cells, and smooth muscle cells. Human TNF-alpha consisits of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 177 aa extracellular domain (ECD). Within the ECD, human TNF-alpha shares 97% aa sequence identity with rhesus and 71%-92% with bovine, canine, cotton rat, equine, feline, mouse, porcine, and rat TNF-alpha. TNF-alpha is produced by a wide variety of immune, epithelial, endothelial, and tumor cells. TNF-alpha is assembled intracellularly to form a noncovalently linked homotrimer which is expressed on the cell surface. Cell surface TNF-alpha can induce the lysis of neighboring tumor cells and virus infected cells, and it can generate its own downstream cell signaling following ligation by soluble TNFR I. Shedding of membrane bound TNF-alpha by TACE/ADAM17 releases the bioactive cytokine, a 55 kDa molecular weight soluble trimer of the TNF-alpha extracellular domain. TNF-alpha binds the ubiquitous 55-60 kDa TNF RI and the hematopoietic cell-restricted 80 kDa TNF RII, both of which are also expressed as homotrimers present on virtually all cell types. Both type I and type II receptors bind TNF-alpha with comparable affinity, although only TNF RI contains a cytoplasmic death domain which triggers the activation of apoptosis. Soluble forms of both types of receptors are released and can neutralize the biological activity of TNF-alpha.

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.

Publications for TNF-alpha Antibody (MAB610)(38)

Publications using MAB610

• A Stumpf, T Brandstett, J Hübner, J Rühe Hydrogel based protein biochip for parallel detection of biomarkers for diagnosis of a Systemic Inflammatory Response Syndrome (SIRS) in human serum PLoS ONE, 2019;14(12):e0225525. 2019 [PMID: 31790441] (ELISA Capture, Human)
• W Sun, N Meednu, A Rosenberg, J Rangel-Mor, V Wang, J Glanzman, T Owen, X Zhou, H Zhang, BF Boyce, JH Anolik, L Xing B cells inhibit bone formation in rheumatoid arthritis by suppressing osteoblast differentiation Nat Commun, 2018;9(1):5127. 2018 [PMID: 30510188] (Neutralization, Mouse)
• Y Wang, J Liu, X Chen, H Sun, S Peng, Y Kuang, J Pi, T Zhuang, L Zhang, Z Yu, B Tomlinson, P Chan, Y Chen, Y Zhang, Y Li Dysfunctional endothelial-derived microparticles promote inflammatory macrophage formation via NF-?B and IL-1? signal pathways J. Cell. Mol. Med., 2018;0(0):. 2018 [PMID: 30334371] (Western Blot, Human)
• S Urra, MC Fischer, JR Martínez, L Véliz, P Orellana, A Solar, K Bohmwald, A Kalergis, C Riedel, AH Corvalán, JC Roa, R Fuentealba, CJ Cáceres, M López-Last, A León, N Droppelman, HE González Differential expression profile of CXCR3 splicing variants is associated with thyroid neoplasia. Potential role in papillary thyroid carcinoma oncogenesis? Oncotarget, 2018;9(2):2445-2467. 2018 [PMID: 29416784] (Western Blot, Human)
• K Ouhara, S Munenaga, M Kajiya, K Takeda, S Matsuda, Y Sato, Y Hamamoto, T Iwata, S Yamasaki, K Akutagawa, N Mizuno, T Fujita, E Sugiyama, H Kurihara The induced RNA-binding protein, HuR targets 3&#039;-UTR region of IL-6 mRNA and enhances its stabilization in periodontitis Clin. Exp. Immunol., 2018;0(0):. 2018 [PMID: 29393507] (Neutralization, Human)
• T Regan, AC Gill, SM Clohisey, MW Barnett, CM Pariante, NA Harrison,, DA Hume, ET Bullmore, TC Freeman Effects of anti-inflammatory drugs on the expression of tryptophan-metabolism genes by human macrophages J. Leukoc. Biol., 2018;103(4):681-692. 2018 [PMID: 29377288] (Bioassay, Human)
• CM Henry, SJ Martin Caspase-8 Acts in a Non-enzymatic Role as a Scaffold for Assembly of a Pro-inflammatory FADDosome Complex upon TRAIL Stimulation Mol. Cell, 2017;65(4):715-729.e5. 2017 [PMID: 28212752] (ICC, Human)
• MC Tanzer, N Khan, JA Rickard, N Etemadi, N Lalaoui, SK Spall, JM Hildebrand, D Segal, M Miasari, D Chau, WL Wong, M McKinlay, SK Chunduru, CA Benetatos, SM Condon, JE Vince, MJ Herold, J Silke Combination of IAP antagonist and IFN? activates novel caspase-10- and RIPK1-dependent cell death pathways Cell Death Differ, 2017;0(0):. 2017 [PMID: 28106882] (Neutralization, Human)
• H Jiang, X Zhang, H Lin Lysine fatty acylation promotes lysosomal targeting of TNF-? Sci Rep, 2016;6(0):24371. 2016 [PMID: 27079798] (IHC-Fr, Human)
• Matt P, Lindqvist U, Kleinau S Elevated Membrane and Soluble CD64: A Novel Marker Reflecting Altered FcgammaR Function and Disease in Early Rheumatoid Arthritis That Can Be Regulated by Anti-Rheumatic Treatment. PLoS ONE, 2015;10(9):e0137474. 2015 [PMID: 26406605] (ELISA Development (Capture), Human)
• Zuo J, Shan Z, Zhou L, Yu J, Liu X, Gao Y Increased CD160 expression on circulating natural killer cells in atherogenesis. J Transl Med, 2015;13(0):188. 2015 [PMID: 26071079] (Neutralization, Human)
• Tudor C, Bruns H, Daniel C, Distel L, Hartmann A, Gerbitz A, Buettner M Macrophages and dendritic cells as actors in the immune reaction of classical Hodgkin lymphoma. PLoS ONE, 2015;9(12):e114345. 2015 [PMID: 25470820] (Neutralization, Human)
• Todd J, Simpson P, Estis J, Torres V, Wub A Reference range and short- and long-term biological variation of interleukin (IL)-6, IL-17A and tissue necrosis factor-alpha using high sensitivity assays. Cytokine, 2013;64(3):660-5. 2013 [PMID: 24128872] (ELISA Development, Human)
• Krepler C, Chunduru S, Halloran M, He X, Xiao M, Vultur A, Villanueva J, Mitsuuchi Y, Neiman E, Benetatos C, Nathanson K, Amaravadi R, Pehamberger H, McKinlay M, Herlyn M The novel SMAC mimetic birinapant exhibits potent activity against human melanoma cells. Clin Cancer Res, 2013;19(7):1784-94. 2013 [PMID: 23403634] (Neutralization, Human)
• Kulbe H, Chakravarty P, Leinster DA, Charles KA, Kwong J, Thompson RG, Coward JI, Schioppa T, Robinson SC, Gallagher WM, Galletta L, Salako MA, Smyth JF, Hagemann T, Brennan DJ, Bowtell DD, Balkwill FR A dynamic inflammatory cytokine network in the human ovarian cancer microenvironment. Cancer Res., 2012;72(1):66-75. 2012 [PMID: 22065722] (IHC-P, Human)
• Liu F, Chen HY, Huang XR, Chung AC, Zhou L, Fu P, Szalai AJ, Lan HY C-reactive protein promotes diabetic kidney disease in a mouse model of type 1 diabetes. Diabetologia, 2011;54(10):2713-23. 2011 [PMID: 21744073] (ELISA Development, Human)
• Zepp JA, Nold-Petry CA, Dinarello CA, Nold MF Protection from RNA and DNA viruses by IL-32. J. Immunol., 2011;186(7):4110-8. 2011 [PMID: 21346229] (Electrochemiluminescent Assay, Human)
• Wu W, Mehta H, Chakrabarty K, Booth JL, Duggan ES, Patel KB, Ballard JD, Coggeshall KM, Metcalf JP Resistance of human alveolar macrophages to Bacillus anthracis lethal toxin. J. Immunol., 2009;183(9):5799-806. 2009 [PMID: 19812208] (ELISA Development, Human)
• Hu H, Kwun J, Aizenstein BD, Knechtle SJ Noninvasive detection of acute and chronic injuries in human renal transplant by elevation of multiple cytokines/chemokines in urine. Transplantation, 2009;87(12):1814-20. 2009 [PMID: 19543058] (Antibody Array Development, Human)
• Gonzalez RM, Seurynck-Servoss SL, Crowley SA Development and validation of sandwich ELISA microarrays with minimal assay interference. J. Proteome Res., 2008;7(6):2406-14. 2008 [PMID: 18422355] (ELISA Microarray Development, Human)
• Biancotto A, Iglehart SJ, Lisco A, Vanpouille C, Grivel JC, Lurain NS, Reichelderfer PS, Margolis LB Upregulation of human cytomegalovirus by HIV type 1 in human lymphoid tissue ex vivo. AIDS Res. Hum. Retroviruses, 2008;24(3):453-62. 2008 [PMID: 18327985] (Luminex Development, Human)
• Kim HG, Kim NR, Gim MG, Lee JM, Lee SY, Ko MY, Kim JY, Han SH, Chung DK Lipoteichoic acid isolated from Lactobacillus plantarum inhibits lipopolysaccharide-induced TNF-alpha production in THP-1 cells and endotoxin shock in mice. J. Immunol., 2008;180(4):2553-61. 2008 [PMID: 18250466] (ELISA Development, Human)
• Qui H, Ferrell EP, Nolan N, Phelps BH, Tabibiazar R, Whitney DH, Naelfski EA Fluorescence single-molecule counting assays for high-sensitivity detection of cytokines and chemokines. Clin. Chem., 2007;53(11):2010-2. 2007 [PMID: 18030696] (ELISA Development, Human)
• John T, Muller RD, Oberholzer A, Zreiqat H, Kohl B, Ertel W, Hostmann A, Tschoeke SK, Schulze-Tanzil G Interleukin-10 modulates pro-apoptotic effects of TNF-alpha in human articular chondrocytes in vitro. Cytokine, 2007;40(3):226-34. 2007 [PMID: 18023359] (ICC, Human)
• Harrison ML, Obermueller E, Maisey NR, Hoare S, Edmonds K, Li NF, Chao D, Hall K, Lee C, Timotheadou E, Charles K, Ahern R, King DM, Eisen T, Corringham R, DeWitte M, Balkwill F, Gore M Tumor necrosis factor alpha as a new target for renal cell carcinoma: two sequential phase II trials of infliximab at standard and high dose. J. Clin. Oncol., 2007;25(29):4542-9. 2007 [PMID: 17925549] (IHC-Fr, Human)
• Todd J, Freese B, Lu A, Held D, Morey J, Livingston R, Goix P Ultrasensitive flow-based immunoassays using single-molecule counting. Clin. Chem., 2007;53(11):1990-5. 2007 [PMID: 17890441] (ELISA Development, Human)
• Grivel JC, Elliott J, Lisco A, Biancotto A, Condack C, Shattock RJ, McGowan I, Margolis L, Anton P HIV-1 pathogenesis differs in rectosigmoid and tonsillar tissues infected ex vivo with CCR5- and CXCR4-tropic HIV-1. AIDS, 2007;21(10):1263-72. 2007 [PMID: 17545702] (Luminex Development, Human)
• Colucci S, Brunetti G, Cantatore FP, Oranger A, Mori G, Quarta L, Cirulli N, Mancini L, Corrado A, Grassi FR, Grano M Lymphocytes and synovial fluid fibroblasts support osteoclastogenesis through RANKL, TNFalpha, and IL-7 in an in vitro model derived from human psoriatic arthritis. J. Pathol., 2007;212(1):47-55. 2007 [PMID: 17370327] (Neutralization, Human)
• Biancotto A, Grivel JC, Iglehart SJ, Vanpouille C, Lisco A, Sieg SF, Debernardo R, Garate K, Rodriguez B, Margolis LB, Lederman MM Abnormal activation and cytokine spectra in lymph nodes of people chronically infected with HIV-1. Blood, 2007;109(10):4272-9. 2007 [PMID: 17289812] (Luminex Development, Human)
• Du Z, Kelly E, Mecklenbrauker I, Agle L, Herrero C, Paik P, Ivashkiv LB Selective regulation of IL-10 signaling and function by zymosan. J. Immunol., 2006;176(8):4785-92. 2006 [PMID: 16585572] (ELISA Development, Human)
• Wunder DM, Mueller MD, Birkhauser MH, Bersinger NA Increased ENA-78 in the follicular fluid of patients with endometriosis. Acta Obstet Gynecol Scand, 2006;85(3):336-42. 2006 [PMID: 16553183] (ELISA Development, Human)
• Hangai M, He S, Hoffmann S, Lim JI, Ryan SJ, Hinton DR Sequential induction of angiogenic growth factors by TNF-alpha in choroidal endothelial cells. J. Neuroimmunol., 2005;171(1):45-56. 2005 [PMID: 16288810] (IHC, Human)
• Mozaffarian D, Rimm EB, King IB, Lawler RL, McDonald GB, Levy WC trans fatty acids and systemic inflammation in heart failure. Am. J. Clin. Nutr., 2004;80(6):1521-5. 2004 [PMID: 15585763] (ELISA Development, Human)
• Phillips TM Rapid analysis of inflammatory cytokines in cerebrospinal fluid using chip-based immunoaffinity electrophoresis. Electrophoresis, 2004;25(10):1652-9. 2004 [PMID: 15188254] (Affinity Chromatography, Human)
• Sharif MN, Tassiulas I, Hu Y, Mecklenbrauker I, Tarakhovsky A, Ivashkiv LB IFN-alpha priming results in a gain of proinflammatory function by IL-10: implications for systemic lupus erythematosus pathogenesis. J. Immunol., 2004;172(10):6476-81. 2004 [PMID: 15128840] (ELISA Development, Human)
• Buch MH, Conaghan PG, Quinn MA, Bingham SJ, Veale D, Emery P True infliximab resistance in rheumatoid arthritis: a role for lymphotoxin alpha? Ann. Rheum. Dis., 2004;63(10):1344-6. 2004 [PMID: 15033655] (IHC-Fr, Human)
• Copeland S, Siddiqui J, Remick D Direct comparison of traditional ELISAs and membrane protein arrays for detection and quantification of human cytokines. J. Immunol. Methods, 2004;284(1):99-106. 2004 [PMID: 14736420] (Array Development, Human)
• Herrero C, Hu X, Li WP, Samuels S, Sharif MN, Kotenko S, Ivashkiv LB Reprogramming of IL-10 activity and signaling by IFN-gamma. J. Immunol., 2003;171(10):5034-41. 2003 [PMID: 14607900] (ELISA Development, Human)

Review for TNF-alpha Antibody (MAB610) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Anonymous WB Human 04/20/2017

Summary

• Application: Western Blot
• Sample Tested: Human breast cancer cell lysate (MCF-7)
• Species: Human
• Lot: AVR1716011

Comments

• Comments: MCF-7 cell extract probed with 1 ug/mL of Mouse anti-Human TNF-alpha Antibody (#MAB610) showing a specific band at 18 kDa. Membrane blocked with 5%BSA in PBS. Buffer: 5% BSA in PBS. Secondary Ab: anti-Mouse IgG 1:5,000.

FAQs for TNF-alpha Antibody (MAB610). (Showing 1 - 4 of 4 FAQs).

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
• Uniprot:
  • P01375()