Rat TNF-alpha Antibody

(7 citations)
(1 Review)
  
  • Species Reactivity
    Rat
  • Specificity
    Detects rat TNF-alpha in ELISAs and Western blots. In ELISAs, this antibody shows less than 3% cross-reactivity with recombinant mouse (rm) TNF‑ alpha and less than 0.2% cross-reactivity with rhTNF‑ alpha, rpTNF‑ alpha, and rhTNF‑ beta. In Western blots, this antibody shows 100% cross-reactivity with rmTNF‑ alpha  and no cross-reactivity with rfeTNF‑ alpha.
  • Source
    Monoclonal Mouse IgG1 Clone # 45418
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant rat TNF-alpha
    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
    1 µg/mL
    Recombinant Rat TNF‑ alpha (Catalog # 510-RT)
    • Rat TNF-alpha Sandwich Immunoassay
      Reagent
  • ELISA Capture (Matched Antibody Pair)
    2-8 µg/mL 
    Rat TNF‑ alpha Antibody (Catalog # MAB510)
  • ELISA Detection (Matched Antibody Pair)
    0.1-0.4 µg/mL 
    Rat TNF‑ alpha Biotinylated Antibody (Catalog # BAF510)
  • ELISA Standard
     
    Recombinant Rat TNF-alpha Protein (Catalog # 510-RT)
  • 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 10-40 µg/mL in the presence of 0.025 ng/mL Recombinant Rat TNF‑ alpha.
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 crystal violet staining. Cytotoxicity elicited by Recombinant Rat TNF‑ alpha (0.025 ng/mL) is neutralized (green line) by increasing concentrations of Rat TNF‑ alpha Monoclonal Antibody (Catalog # MAB510). The ND50 is typically 10-40 µg/mL in the presence of the metabolic inhibitor actinomycin D (1 µg/mL).
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 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, 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 macaque 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.

7 Citations: Showing 1 - 7
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Applications
Sample Type
  1. Urban traffic-derived nanoparticulate matter reduces neurite outgrowth via TNFalpha in vitro.
    Authors: Cheng H, Davis D, Hasheminassab S, Sioutas C, Morgan T, Finch C
    J Neuroinflammation, 2016;13(1):19.
    Species: Rat
    Sample Type: Whole Cells
    Application: Neut
  2. Beneficial Effect of Moderate Exercise in Kidney of Rat after Chronic Consumption of Cola Drinks
    Authors: G Cao, J González, A Müller, G Ottaviano, G Ambrosio, JE Toblli, J Milei
    PLoS ONE, 2016;11(3):e0152461.
    Species: Rat
    Sample Type: Whole Tissue
    Application: IHC - Paraffin embedded
  3. Neuronal synapse formation induced by microglia and interleukin 10.
    Authors: Lim S, Park E, You B, Jung Y, Park A, Park S, Lee J
    PLoS ONE, 2013;8(11):e81218.
    Species: Rat
    Sample Type: Whole Cells
    Application: Neut
  4. Dose-dependent changes in neuroinflammatory and arachidonic acid cascade markers with synaptic marker loss in rat lipopolysaccharide infusion model of neuroinflammation.
    BMC Neurosci, 2012;13(0):50.
    Species: Rat
    Sample Type: Tissue Homogenates
    Application: WB
  5. Rat pneumonia and soft-tissue infection models for the study of Acinetobacter baumannii biology.
    Authors: Russo TA, Beanan JM, Olson R, MacDonald U, Luke NR, Gill SR, Campagnari AA
    Infect. Immun., 2008;76(8):3577-86.
    Species: Rat
    Sample Type: BALF
    Application: ELISA Development
  6. In vivo modulation of LPS-induced alterations in brain and peripheral cytokines and HPA axis activity by cannabinoids.
    Authors: Roche M, Diamond M, Finn DP
    J. Neuroimmunol., 2006;181(1):57-67.
    Species: Rat
    Sample Type: Plasma
    Application: ELISA Development
  7. Oxidative mitochondrial DNA damage and deletion in hepatocytes of rejecting liver allografts in rats: role of TNF-alpha.
    Authors: Nagakawa Y, Williams GM, Zheng Q, Tsuchida A, Aoki T, Montgomery RA, Klein AS, Sun Z
    Hepatology, 2005;42(1):208-15.
    Species: Rat
    Sample Type: Whole Tissue
    Application: IHC
Isotype Controls
Description Application Cat# Citations Images  

Mouse IgG1 Isotype Control

Ctrl MAB002 138  
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Mouse IgG1 Isotype Control

Ctrl MAB002R 1
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Primary Antibodies
Description Application Cat# Citations Images  

Rat TNF-alpha Antibody

Neut MAB510R  
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Secondary Antibodies
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Mouse IgG HRP-conjugated Antibody

WB HAF007 26  
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Mouse F(ab)2 IgG (H+L) PE-conjugated Antibody

Flow F0102B 10  
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Donkey Anti-Mouse IgG NorthernLights™ NL557-conjugate Antibody

Flow, IHC, ICC NL007 9
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Donkey Anti-Mouse IgG Biotinylated Antibody

WB BAF018 1
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Mouse IgG HRP-conjugated Antibody

WB HAF018 9  
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Donkey Anti-Mouse IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL008 8
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Mouse F(ab)2 IgG (H+L) APC-conjugated Antibody

Flow F0101B 2  
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Donkey Anti-Mouse IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL009 5
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Goat Anti-Mouse IgG Biotinylated Antibody

WB BAF007 4
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Mouse F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

Flow F0103B 5  
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Mouse IgG Antibody

WB AF007 4
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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
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Mouse IgG VisUCyte HRP Polymer Antibody

IHC VC001  
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Goat Anti-Mouse IgG Fc Affinity Purified Polyclonal Ab

G-202-C 2
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Mouse F(ab)2 IgG (H+L) PerCP-conjugated Antibody

Flow F0114  
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ELISA Rat TNF-alpha  Antibody MAB510
ELISA: Rat TNF-alpha Antibody [MAB510]

Summary

ApplicationELISA
Sample TestedSerum and Plasma

Other Experimental Details

Other Experimental DetailsAn ELISA was build targeting rat TNFa using this antibody as a capture and BAF510 as the detection. The assay was used to measure TNFa in rat serum and plasma samples.

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