Rat TNF-alpha Antibody

(17 citations)   
  • Species Reactivity
    Rat
  • 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
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant rat TNF-alpha
    Leu80-Leu235
    Accession # P16599
  • Formulation
    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.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    Recombinant Rat TNF‑ alpha (Catalog # 510-RT)
  • Immunocytochemistry
    5-15 µg/mL
    See below
  • Neutralization
    Measured by its ability to neutralize TNF‑ alpha -induced cytotoxicity in the L‑929 mouse fibroblast cell line. Matthews, N. and M. L. Neale (1987) in Lymphokines and Interferons, A Practical Approach. Clemens, M. J. et al. (eds): IRL Press. 221. The Neutralization Dose (ND50) is typically 0.15-0.75 µg/mL in the presence of 0.25 ng/mL Recombinant Rat TNF‑ alpha and 1 µg/mL actinomycin D.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Cytotoxicity Induced by TNF‑ alpha and Neutralization by Rat TNF‑ alpha Antibody. 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).
Immunocytochemistry
TNF‑ alpha in Rat Splenocytes. 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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & 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.
Background: TNF-alpha

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).

  • References:
    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.
  • Long Name:
    Tumor Necrosis Factor alpha
  • Entrez Gene IDs:
    7124 (Human); 21926 (Mouse); 24835 (Rat); 397086 (Porcine); 280943 (Bovine); 403922 (Canine); 100033834 (Equine); 493755 (Feline); 100009088 (Rabbit)
  • Alternate Names:
    APC1 protein; Cachectin; Cachetin; DIF; TNF; TNF, monocyte-derived; tnfa; tnf-a; TNFalpha; TNF-alpha; TNF-alphacachectin; TNFATNF, macrophage-derived; 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
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

17 Citations: Showing 1 - 10
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Species
Applications
Sample Type
  1. Effects of TNF-? on penile structure alteration in rats with hyperprolactinemia
    Authors: ZL Wang, LY Yang, HH Chen, HH Lin, YT Tsai, WJ Huang
    PLoS ONE, 2017;12(8):e0181952.
    Species: Rat
    Sample Type: In Vivo
    Application: Neut
  2. Prolonged neuroinflammation after lipopolysaccharide exposure in aged rats.
    Authors: Fu H, Yang T, Xiao W, Fan L, Wu Y, Terrando N, Wang T
    PLoS ONE, 2014;9(8):e106331.
    Species: Rat
    Sample Type: Whole Tissue
    Application: IHC Frozen
  3. Time-dependent effects of training on cardiovascular control in spontaneously hypertensive rats: role for brain oxidative stress and inflammation and baroreflex sensitivity.
    Authors: Masson G, Costa T, Yshii L, Fernandes D, Soares P, Laurindo F, Scavone C, Michelini L
    PLoS ONE, 2014;9(5):e94927.
    Species: Rat
    Sample Type: Whole Tissue
    Application: IHC Frozen
  4. Microglial cells contribute to endogenous brain defenses after acute neonatal focal stroke.
    Authors: Faustino JV, Wang X, Johnson CE
    J. Neurosci., 2011;31(36):12992-3001.
    Species: Rat
    Sample Type: Whole Tissue
    Application: IHC
  5. Adenovirus-Mediated Kallistatin Gene Transfer Ameliorates Disease Progression in a Rat Model of Osteoarthritis Induced by Anterior Cruciate Ligament Transection.
    Authors: Hsieh J, Shen PC, Shiau AL, Wu CL, Lee CH, Jou IM, Wang CR, Chao J, Chao L, Teo ML
    2009;20(0):147.
    Species: Rat
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  6. Mechanisms underlying the nociceptive responses induced by platelet-activating factor (PAF) in the rat paw.
    Authors: Marotta DM, Costa R, Motta EM, Fernandes ES, Medeiros R, Quintao NL, Campos MM, Calixto JB
    Biochem. Pharmacol., 2009;77(7):1223-35.
    Species: Rat
    Sample Type: In Vivo
    Application: In vivo
  7. Distribution and tumor necrosis factor-alpha isoform binding specificity of locally administered etanercept into injured and uninjured rat sciatic nerve.
    Authors: Kato K, Kikuchi S, Shubayev VI, Myers RR
    Neuroscience, 2009;160(2):492-500.
    Species: Rat
    Sample Type: Tissue Homogenates
    Application: WB
  8. Tumor necrosis factor alpha: activity dependent expression and promotion of cortical column sleep in rats.
    Authors: Churchill L, Rector DM, Yasuda K, Fix C, Rojas MJ, Yasuda T, Krueger JM
    Neuroscience, 2008;156(1):71-80.
    Species: Rat
    Sample Type: Tissue Homogenates
    Application: WB
  9. Supraspinal glial-neuronal interactions contribute to descending pain facilitation.
    Authors: Wei F, Guo W, Zou S, Ren K, Dubner R
    J. Neurosci., 2008;28(42):10482-95.
    Species: Rat
    Sample Type: Tissue Homogenates
    Application: WB
  10. Deleterious role of TNF-alpha in retinal ischemia-reperfusion injury.
    Authors: Berger S, Savitz SI, Nijhawan S, Singh M, David J, Rosenbaum PS, Rosenbaum DM
    Invest. Ophthalmol. Vis. Sci., 2008;49(8):3605-10.
    Species: Rat
    Sample Type: In Vivo
    Application: In vivo
  11. Non-steroidal anti-inflammatory drug-induced small intestinal damage is Toll-like receptor 4 dependent.
    Authors: 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
    Gut, 2007;57(2):181-7.
    Species: Rat
    Sample Type: In Vivo
    Application: In vivo
  12. TNFalpha and TNF receptor 1 expression in the mixed neuronal-glial cultures of hippocampal dentate gyrus exposed to glutamate or trimethyltin.
    Authors: Figiel I, Dzwonek K
    Brain Res., 2007;1131(1):17-28.
    Species: Rat
    Sample Type: Whole Cells
    Application: ICC
  13. Wnt-induced secreted protein-1 is a prohypertrophic and profibrotic growth factor.
    Authors: Colston JT, de la Rosa SD, Koehler M, Gonzales K, Mestril R, Freeman GL, Bailey SR, Chandrasekar B
    Am. J. Physiol. Heart Circ. Physiol., 2007;293(3):H1839-46.
    Species: Rat
    Sample Type: Whole Cells
    Application: Neut
  14. Thrombin induces neurodegeneration and microglial activation in the cortex in vivo and in vitro: proteolytic and non-proteolytic actions.
    Authors: Lee da Y, Park KW, Jin BK
    Biochem. Biophys. Res. Commun., 2006;346(3):727-38.
    Species: Rat
    Sample Type: Whole Tissue
    Application: IHC
  15. TNFalpha-induced AMPA-receptor trafficking in CNS neurons; relevance to excitotoxicity?
    Authors: Leonoudakis D, Braithwaite SP, Beattie MS, Beattie EC
    Neuron Glia Biol., 2004;1(3):263-273.
    Species: Rat
    Sample Type: Whole Cells
    Application: ICC
  16. Differential cytokine response in interstitial fluid in skin and serum during experimental inflammation in rats.
    Authors: Nedrebo T, Reed RK, Jonsson R, Berg A, Wiig H
    J. Physiol. (Lond.), 2004;556(0):193-202.
    Species: Rat
    Sample Type: Whole Tissue
    Application: IHC Frozen
  17. Glucocorticoids play a fundamental role in protecting the brain during innate immune response.
    Authors: Nadeau S, Rivest S
    J. Neurosci., 2003;23(13):5536-44.
    Species: Rat
    Sample Type:
    Application: Neut
Expand to show all 17 Citations
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