TNF-alpha Antibody [Unconjugated]

Recombinant Human TNF‑ alpha (Catalog # 210-TA) induces cytotoxicity in the L‑929 mouse fibroblast cell line in a dose-dependent manner (orange line). Cytotoxicity elicited by Recombinant Human TNF‑ alpha (1.5 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human TNF‑ alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-210-NA). The ND50 is typically 0.01‑0.06 μg/mL.

TNF‑ alpha was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Goat Anti-Human TNF‑ alpha Antigen Affinity-purified Polyclonal Antibody (Catalog #AF-210-NA) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

• Reactivity: Hu
• Applications: WB, B/N, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

TNF-alpha Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human TNF-alpha
  Val77-Leu233
  Accession # P01375
• Specificity: Detects human TNF‑ alpha in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: TNF
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Western Blot 0.1 ug/mL
  • Immunocytochemistry 5-15 ug/mL
  • Neutralization 0.01-0.06 ug/mL

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
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 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 [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 (AF-210-NA)(23)

Publications using AF-210-NA

• K Ni, A Gill, D Cao, K Koike, KS Schweitzer, S Garantziot, I Petrache Intravascular heavy chain-modification of hyaluronan during endotoxic shock Biochem Biophys Rep, 2019;17(0):114-121. 2019 [PMID: 30623115] (Western Blot, Human)
• M Kim, S Pyo, CH Kang, CO Lee, HK Lee, SU Choi, CH Park Folate receptor 1 (FOLR1) targeted chimeric antigen receptor (CAR) T cells for the treatment of gastric cancer PLoS ONE, 2018;13(6):e0198347. 2018 [PMID: 29874279] (Neutralization, Human)
• V Rothhammer, DM Borucki, JE Kenison, P Hewson, Z Wang, R Bakshi, DH Sherr, FJ Quintana Detection of aryl hydrocarbon receptor agonists in human samples Sci Rep, 2018;8(1):4970. 2018 [PMID: 29563571] (Neutralization, Human)
• R Jalili, J Horecka, JR Swartz, RW Davis, HHJ Persson Streamlined circular proximity ligation assay provides high stringency and compatibility with low-affinity antibodies Proc. Natl. Acad. Sci. U.S.A., 2018;0(0):. 2018 [PMID: 29339495] (Affinity Assay)
• M A Rochat Promising Role of Toll-Like Receptor 8 Agonist in Concert with Prostratin for Activation of Silent HIV J. Virol, 2016;0(0):. 2016 [PMID: 27928016] (Neutralization, Human)
• Inflammatory Adipokines Decrease Expression of Two High Molecular Weight Isoforms of Tropomyosin Similar to the Change in Type 2 Diabetic Patients PLoS ONE, 2016;11(9):e0162908. 2016 [PMID: 27649540] (Bioassay, Human)
• Holmes T, Wilson E, Black E, Benest A, Vaz C, Tan B, Tanavde V, Cook G Licensed human natural killer cells aid dendritic cell maturation via TNFSF14/LIGHT. Proc Natl Acad Sci U S A, 2014;111(52):E5688-96. 2014 [PMID: 25512551] (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)
• Wang R, Jaw JJ, Jaw J, Stutzman N, Stutzman NC, Zou Z, Sun PD Natural killer cell-produced IFN-gamma and TNF-alpha induce target cell cytolysis through up-regulation of ICAM-1. J Leukoc Biol, 0;91(2):299-309. 0 [PMID: 22045868] (Neutralization, Human)
• Cunin P, Caillon A, Corvaisier M, Garo E, Scotet M, Blanchard S, Delneste Y, Jeannin P The tachykinins substance P and hemokinin-1 favor the generation of human memory Th17 cells by inducing IL-1beta, IL-23, and TNF-like 1A expression by monocytes. J. Immunol., 2011;186(7):4175-82. 2011 [PMID: 21368235] (Neutralization, Human)
• Hasegawa H, Thomas HJ, Schooley K, Born TL Native IL-32 is released from intestinal epithelial cells via a non-classical secretory pathway as a membrane-associated protein. Cytokine, 2011;53(1):74-83. 2011 [PMID: 20926308] (Western Blot, Human)
• Guo PF, Du MR, Wu HX, Lin Y, Jin LP, Li DJ Thymic stromal lymphopoietin from trophoblasts induces dendritic cell-mediated regulatory TH2 bias in the decidua during early gestation in humans. Blood, 2010;116(12):2061-9. 2010 [PMID: 20538796] (IHC, Human)
• Smith AM, Rahman FZ, Hayee B, Graham SJ, Marks DJ, Sewell GW, Palmer CD, Wilde J, Foxwell BM, Gloger IS, Sweeting T, Marsh M, Walker AP, Bloom SL, Segal AW Disordered macrophage cytokine secretion underlies impaired acute inflammation and bacterial clearance in Crohn's disease. J. Exp. Med., 2009;206(9):1883-97. 2009 [PMID: 19652016] (Western Blot, Human)
• Navenot JM, Fujii N, Peiper SC KiSS1 metastasis suppressor gene product induces suppression of tyrosine kinase receptor signaling to Akt, tumor necrosis factor family ligand expression, and apoptosis. Mol. Pharmacol., 2009;75(5):1074-83. 2009 [PMID: 19201817] (Neutralization, Human)
• Skibinski G, Elborn JS, Ennis M Bronchial epithelial cell growth regulation in fibroblast cocultures: the role of hepatocyte growth factor. Am. J. Physiol. Lung Cell Mol. Physiol., 2007;293(1):L69-76. 2007 [PMID: 17384084] (Neutralization, Human)
• Lee SY, Cherla RP, Tesh VL Simultaneous induction of apoptotic and survival signaling pathways in macrophage-like THP-1 cells by Shiga toxin 1. Infect. Immun., 2007;75(3):1291-302. 2007 [PMID: 17194804] (Functional Assay, Human)
• Hunter I, Nixon GF Spatial compartmentalization of tumor necrosis factor (TNF) receptor 1-dependent signaling pathways in human airway smooth muscle cells. Lipid rafts are essential for TNF-alpha-mediated activation of RhoA but dispensable for the activation of the NF-kappaB and MAPK pathways. J. Biol. Chem., 2006;281(45):34705-15. 2006 [PMID: 16982613] (Neutralization, Human)
• Stearns-Kurosawa DJ, Lupu F, Taylor FB, Kinasewitz G, Kurosawa S Sepsis and pathophysiology of anthrax in a nonhuman primate model. Am. J. Pathol., 2006;169(2):433-44. 2006 [PMID: 16877346] (ELISA Development, Primate - Papio cynocephalus (Yellow Baboon))
• Brando C, Mukhopadhyay S, Kovacs E, Medina R, Patel P, Catina TL, Campbell KS, Santoli D Receptors and lytic mediators regulating anti-tumor activity by the leukemic killer T cell line TALL-104. J. Leukoc. Biol., 2005;78(2):359-71. 2005 [PMID: 15937142] (Neutralization, Human)
• Andersson By U, Tani E, Andersson U Tumor necrosis factor, interleukin 11, and leukemia inhibitory factor produced by Langerhans cells in Langerhans cell histiocytosis. J. Pediatr. Hematol. Oncol., 2004;26(11):706-11. 2004 [PMID: 15543003] (ICC, Human)
• Shahbakhti H, Watson RE, Azurdia RM, Ferreira CZ, Garmyn M, Rhodes LE Influence of eicosapentaenoic acid, an omega-3 fatty acid, on ultraviolet-B generation of prostaglandin-E2 and proinflammatory cytokines interleukin-1 beta, tumor necrosis factor-alpha, interleukin-6 and interleukin-8 in human skin in vivo. Photochem. Photobiol., 2004;80(2):231-5. 2004 [PMID: 15362934] (IHC, Human)
• Park JS, Kim YS, Jee YK, Myong NH, Lee KY Interleukin-8 production in tuberculous pleurisy: role of mesothelial cells stimulated by cytokine network involving tumour necrosis factor-alpha and interleukin-1 beta. Scand. J. Immunol., 2003;57(5):463-9. 2003 [PMID: 12753503] (Neutralization, Human)
• Edgeworth JD, Spencer J, Phalipon A, Griffin GE, Sansonetti PJ Cytotoxicity and interleukin-1beta processing following Shigella flexneri infection of human monocyte-derived dendritic cells. Eur. J. Immunol., 2002;32(5):1464-71. 2002 [PMID: 11981835] (Western Blot, Human)

FAQs for TNF-alpha Antibody (AF-210-NA). (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()