TNF-alpha Antibody [Unconjugated]

TNF‑ alpha was detected in immersion fixed rat splenocytes using Goat Anti-Rat TNF‑ alpha Antigen Affinity-purified Polyclonal Antibody(Catalog # AF-510-NA) at 15 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 cytoplasmic. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Recombinant Rat TNF‑ alpha (Catalog # 510-RT) induces cytotoxicity in the the L‑929 mouse fibroblast cell line in a dose-dependent manner (orange line), as measured by Resazurin. Cytotoxicity elicited by Recombinant Rat TNF‑ alpha (0.25 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Rat TNF‑ alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-510-NA). The ND50 is typically 0.15‑0.75 µg/mL in the presence of the metabolic inhibitor actinomycin D (1 µg/mL).

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

TNF-alpha Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant rat TNF-alpha
  Leu80-Leu235
  Accession # P16599
• Specificity: Detects rat TNF‑ alpha in direct ELISAs and Western blots. In Western blots, approximately 25% cross-reactivity with recombinant porcine TNF-alpha is observed and less than 1% cross-reactivity with recombinant human (rh) TNF-beta, rhTNF sRI, and rhTNF sRII is observed. In direct ELISAs, approximately 40% cross-reactivity with recombinant canine TNF-alpha, recombinant feline TNF-alpha, and recombinant rhesus macaque TNF-alpha is observed. Neutralizes the biological activity of rat TNF-alpha. This antibody will also neutralize the biological activity of murine TNF-alpha, but weak neutralization of rhTNF-alpha and no neutralization of rhTNF-beta was observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: TNF
• Purity: Immunogen affinity purified
• Purity Statement: Antigen Affinity-purified
• 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 0.15-0.75 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
• Purity: Immunogen affinity purified
• 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 ) 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, apoptosis, and immune system development. TNF-alpha is produced by a wide variety of immune and epithelial cell types (1, 2). Rat TNF-alpha consisits of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 179 aa extracellular domain (ECD) (3). Within the ECD, rat TNF-alpha shares 94% aa sequence identity with mouse and 69% - 76% with bovine, canine, cotton rat, equine, feline, human, porcine, and rhesus TNF-alpha. The 26 kDa type 2 transmembrane protein is assembled intracellularly to form a noncovalently linked homotrimer (4). Ligation of this complex induces reverse signaling that promotes lymphocyte co-stimulation but diminishes monocyte responsiveness (5). Cleavage of membrane bound TNF-alpha by TACE/ADAM17 releases a 55 kDa soluble trimeric form of TNF-alpha (6, 7). TNF-alpha trimers bind the ubiquitous TNF RI and the hematopoietic cell-restricted TNF RII, both of which are also expressed as homotrimers (1, 8). TNF-alpha regulates lymphoid tissue development through control of apoptosis (2). It also promotes inflammatory responses by inducing the activation of vascular endothelial cells and macrophages (2). TNF-alpha is a key cytokine in the development of several inflammatory disorders (9). It contributes to the development of type 2 diabetes through its effects on insulin resistance and fatty acid metabolism (10, 11).

1. Idriss, H.T. and J.H. Naismith (2000) Microsc. Res. Tech. 50:184.
2. Hehlgans, T. and K. Pfeffer (2005) Immunology 115:1.
3. Estler, H.C. et al. (1992) Biol. Chem. Hoppe-Seyler 373:271.
4. Tang, P. et al. (1996) Biochemistry 35:8216.
5. Eissner G. et al. (2004) Cytokine Growth Factor Rev. 15:353.
6. Black, R.A. et al. (1997) Nature 385:729.
7. Moss, M.L. et al. (1997) Nature 385:733.
8. Loetscher, H. et al. (1991) J. Biol. Chem. 266:18324.
9. Clark, I.A. (2007) Cytokine Growth Factor Rev. 18:335.
10. Romanatto, T. et al. (2007) Peptides 28:1050.
11. Hector, J. et al. (2007) Horm. Metab. Res. 39:250.

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-510-NA)(24)

Publications using AF-510-NA

• ZH Wang, J Xiang, X Liu, SP Yu, FP Manfredsso, IM Sandoval, S Wu, JZ Wang, K Ye Deficiency in BDNF/TrkB Neurotrophic Activity Stimulates δ-Secretase by Upregulating C/EBP? in Alzheimer&#039;s Disease Cell Rep, 2019;28(3):655-669.e5. 2019 [PMID: 31315045] (Neutralization, Rat)
• HC Lai, CH Lu, CS Wong, BF Lin, SM Chan, CY Kuo, ZF Wu Baicalein attenuates neuropathic pain and improves sciatic nerve function recovery in rats with partial sciatic nerve transection J Chin Med Assoc, 2018;0(0):. 2018 [PMID: 30139671] (Rat)
• YW Zhao, YQ Pan, MM Tang, WJ Lin Blocking p38 Signaling Reduces the Activation of Pro-inflammatory Cytokines and the Phosphorylation of p38 in the Habenula and Reverses Depressive-Like Behaviors Induced by Neuroinflammation Front Pharmacol, 2018;9(0):511. 2018 [PMID: 29867510] (Western Blot, Rat)
• L Ma, Y Jiang, Y Dong, J Gao, B Du, D Liu Anti-TNF-alpha antibody attenuates subarachnoid hemorrhage-induced apoptosis in the hypothalamus by inhibiting the activation of Erk Neuropsychiatr Dis Treat, 2018;14(0):525-536. 2018 [PMID: 29497296] (Western Blot, Rat)
• SA Chong, S Balosso, C Vandenplas, G Szczesny, E Hanon, K Claes, X Van Damme, B Danis, J Van Eyll, C Wolff, A Vezzani, RM Kaminski, I Niespodzia Intrinsic Inflammation Is a Potential Anti-Epileptogenic Target in the Organotypic Hippocampal Slice Model Neurotherapeutics, 2018;0(0):. 2018 [PMID: 29464573] (Neutralization, Rat)
• E DeVallance, KW Branyan, K Lemaster, IM Olfert, DM Smith, EE Pistilli, JC Frisbee, PD Chantler Aortic dysfunction in metabolic syndrome mediated by perivascular adipose tissue TNF? and NOX2 dependent pathway Exp. Physiol., 2018;0(0):. 2018 [PMID: 29349831] (Neutralization, Rat)
• ZL Wang, LY Yang, HH Chen, HH Lin, YT Tsai, WJ Huang Effects of TNF-? on penile structure alteration in rats with hyperprolactinemia PLoS ONE, 2017;12(8):e0181952. 2017 [PMID: 28763467] (Neutralization, Rat)
• JS Medel-Matu, A Reynolds, D Shin, R Sankar, A Mazarati Regulation of kindling epileptogenesis by hippocampal Toll-like receptors 2 Epilepsia, 2017;0(0):. 2017 [PMID: 28632301] (IHC, Rat)
• Fu H, Yang T, Xiao W, Fan L, Wu Y, Terrando N, Wang T Prolonged neuroinflammation after lipopolysaccharide exposure in aged rats. PLoS ONE, 2014;9(8):e106331. 2014 [PMID: 25170959] (IHC-Fr, Rat)
• Masson G, Costa T, Yshii L, Fernandes D, Soares P, Laurindo F, Scavone C, Michelini L Time-dependent effects of training on cardiovascular control in spontaneously hypertensive rats: role for brain oxidative stress and inflammation and baroreflex sensitivity. PLoS ONE, 2014;9(5):e94927. 2014 [PMID: 24788542] (IHC-Fr, Rat)
• Faustino JV, Wang X, Johnson CE Microglial cells contribute to endogenous brain defenses after acute neonatal focal stroke. J. Neurosci., 2011;31(36):12992-3001. 2011 [PMID: 21900578] (IHC, Rat)
• Hsieh J, Shen PC, Shiau AL, Wu CL, Lee CH, Jou IM, Wang CR, Chao J, Chao L, Teo ML Adenovirus-Mediated Kallistatin Gene Transfer Ameliorates Disease Progression in a Rat Model of Osteoarthritis Induced by Anterior Cruciate Ligament Transection. 2009;20(0):147. 2009 [PMID: 20377366] (IHC-P, Rat)
• Marotta DM, Costa R, Motta EM, Fernandes ES, Medeiros R, Quintao NL, Campos MM, Calixto JB Mechanisms underlying the nociceptive responses induced by platelet-activating factor (PAF) in the rat paw. Biochem. Pharmacol., 2009;77(7):1223-35. 2009 [PMID: 19283893] (Neutralization, Rat)
• Kato K, Kikuchi S, Shubayev VI, Myers RR Distribution and tumor necrosis factor-alpha isoform binding specificity of locally administered etanercept into injured and uninjured rat sciatic nerve. Neuroscience, 2009;160(2):492-500. 2009 [PMID: 19250961] (Western Blot, Rat)
• Wei F, Guo W, Zou S, Ren K, Dubner R Supraspinal glial-neuronal interactions contribute to descending pain facilitation. J. Neurosci., 2008;28(42):10482-95. 2008 [PMID: 18923025] (Western Blot, Rat)
• Churchill L, Rector DM, Yasuda K, Fix C, Rojas MJ, Yasuda T, Krueger JM Tumor necrosis factor alpha: activity dependent expression and promotion of cortical column sleep in rats. Neuroscience, 2008;156(1):71-80. 2008 [PMID: 18694809] (Western Blot, Rat)
• Berger S, Savitz SI, Nijhawan S, Singh M, David J, Rosenbaum PS, Rosenbaum DM Deleterious role of TNF-alpha in retinal ischemia-reperfusion injury. Invest. Ophthalmol. Vis. Sci., 2008;49(8):3605-10. 2008 [PMID: 18660426] (Neutralization, Rat)
• Watanabe T, Higuchi K, Kobata A, Nishio H, Tanigawa T, Shiba M, Tominaga K, Fujiwara Y, Oshitani N, Asahara T, Nomoto K, Takeuchi K, Arakawa T Non-steroidal anti-inflammatory drug-induced small intestinal damage is Toll-like receptor 4 dependent. Gut, 2007;57(2):181-7. 2007 [PMID: 17639086] (Neutralization, Rat)
• Colston JT, de la Rosa SD, Koehler M, Gonzales K, Mestril R, Freeman GL, Bailey SR, Chandrasekar B Wnt-induced secreted protein-1 is a prohypertrophic and profibrotic growth factor. Am. J. Physiol. Heart Circ. Physiol., 2007;293(3):H1839-46. 2007 [PMID: 17616748] (Neutralization, Rat)
• Figiel I, Dzwonek K TNFalpha and TNF receptor 1 expression in the mixed neuronal-glial cultures of hippocampal dentate gyrus exposed to glutamate or trimethyltin. Brain Res., 2007;1131(1):17-28. 2007 [PMID: 17161388] (ICC, Rat)
• Lee da Y, Park KW, Jin BK Thrombin induces neurodegeneration and microglial activation in the cortex in vivo and in vitro: proteolytic and non-proteolytic actions. Biochem. Biophys. Res. Commun., 2006;346(3):727-38. 2006 [PMID: 16777064] (IHC, Rat)
• Leonoudakis D, Braithwaite SP, Beattie MS, Beattie EC TNFalpha-induced AMPA-receptor trafficking in CNS neurons; relevance to excitotoxicity? Neuron Glia Biol., 2004;1(3):263-273. 2004 [PMID: 16520832] (ICC, Rat)
• Nedrebo T, Reed RK, Jonsson R, Berg A, Wiig H Differential cytokine response in interstitial fluid in skin and serum during experimental inflammation in rats. J. Physiol. (Lond.), 2004;556(0):193-202. 2004 [PMID: 14724215] (IHC-Fr, Rat)
• Nadeau S, Rivest S Glucocorticoids play a fundamental role in protecting the brain during innate immune response. J. Neurosci., 2003;23(13):5536-44. 2003 [PMID: 12843254] (Neutralization, Rat)

FAQs for TNF-alpha Antibody (AF-510-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:
  • P16599()