Porcine TNF-alpha Antibody

(2 citations)   
  • Species Reactivity
    Porcine
  • Specificity
    Detects porcine TNF-alpha in direct ELISAs and Western blots. In direct ELISAs, approximately 13% cross-reactivity with recombinant human (rh) TNF-alpha  is observed and no cross-reactivity with recombinant mouse TNF-alpha, recombinant rat TNF-alpha, rhTNF-beta, rhAPRIL, rhFas Ligand, rhGITR Ligand, rhLIGHT, rhTrance, and rhVEGF is observed.
  • Source
    Monoclonal Mouse IgG1 Clone # 103314
  • Purification
    Protein A or G purified from ascites
  • Immunogen
    E. coli-derived recombinant porcine TNF-alpha
    Arg78-Leu232
    Accession # P23563
  • 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
  • Neutralization
    Measured by its ability to neutralize TNF‑ alpha -induced cytotoxicity in the PK‑15 porcine kidney epithelial 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; Bertoni et al. (1993) J.  Immunol. Meth. 160:267. The Neutralization Dose (ND50) is typically 0.015-0.06 µg/mL in the presence of 0.05 ng/mL Recombinant Porcine 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 Porcine TNF‑ alpha Antibody.
Recombinant Porcine TNF‑ alpha (Catalog # 690‑PT) induces cytotoxicity in the the PK‑15 porcine kidney epithelial cell line in a dose-dependent manner (orange line), as measured by crystal violet staining. Cytotoxicity elicited by Recombinant Porcine TNF‑ alpha (0.05 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Porcine TNF‑ alpha Monoclonal Antibody (Catalog # MAB6901). The ND50 is typically 0.015-0.06 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 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). Porcine TNF-alpha consisits of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 176 aa extracellular domain (ECD) (3). Within the ECD, porcine TNF-alpha shares 69%‑86% aa sequence identity with bovine, canine, cotton rat, equine, feline, human, mouse, rat, 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 costimulation 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. Pauli, U. et al. (1989) Gene 81:185.
    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.

2 Citations: Showing 1 - 2
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Applications
Sample Type
  1. Characterization of a Broadly Reactive Anti-CD40 Agonistic Monoclonal Antibody for Potential Use as an Adjuvant
    Authors: C Martin, SD Waghela, S Lokhandwal, A Ambrus, J Bray, C Vuong, V Vinodkumar, PJ Dominowski, S Rai, D Mwangi, DL Foss, W Mwangi
    PLoS ONE, 2017;12(1):e0170504.
    Species: Porcine
    Sample Type: Whole Cells
    Application: Flow
  2. Enhanced integrative repair of the porcine meniscus in vitro by inhibition of interleukin-1 or tumor necrosis factor alpha.
    Authors: McNulty AL, Moutos FT, Weinberg JB, Guilak F
    Arthritis Rheum., 2007;56(9):3033-42.
    Species: Porcine
    Sample Type: Whole Cells
    Application: Neut
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