Canine TNF-alpha Biotinylated Antibody

  
  • Species Reactivity
    Canine
  • Specificity
    Detects canine TNF-alpha in ELISAs and Western blots. In sandwich immunoassays, approximately 40% cross‑reactivity with recombinant human TNF-alpha and recombinant rhesus macaque TNF-alpha  is observed, less than 2% cross-reactivity with recombinant porcine (rp) TNF-alpha  is observed, and less than 0.5% cross-reactivity with recombinant mouse (rm) TNF-alpha, recombinant rat (rr) TNF-alpha, recombinant cotton rat (rcr) TNF-alpha, and recombinant equine TNF-alpha is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant canine TNF-alpha
    Val77-Leu233
    Accession # P51742
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
  • Label
    Biotin
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    Recombinant Canine TNF‑ alpha (Catalog # 1507-CT)
  • Immunocytochemistry
    5-15 µg/mL
    See below
    • Canine TNF-alpha Sandwich Immunoassay
      Reagent
  • ELISA Capture (Matched Antibody Pair)
    2-8 µg/mL 
    Canine TNF‑ alpha Antibody (Catalog # MAB1507)
  • ELISA Detection (Matched Antibody Pair)
    0.1-0.4 µg/mL 
    Canine TNF‑ alpha Biotinylated Antibody (Catalog # BAF1507)
  • ELISA Standard
     
    Recombinant Canine TNF-alpha Protein (Catalog # 1507-CT)
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
Immunocytochemistry
TNF‑ alpha in Canine PBMCs. TNF‑ alpha was detected in immersion fixed canine peripheral blood mononuclear cells (PBMCs) stimulated with PMA and calcium ionomycin using Goat Anti-Canine TNF‑ alpha Biotinylated Antigen Affinity-purified Polyclonal Antibody (Catalog # BAF1507) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Streptavidin (red; Catalog # NL999) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. 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.
  • 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). Canine TNF-alpha consisits of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 177 aa extracellular domain (ECD) (3). Within the ECD, canine TNF-alpha shares
84‑94% aa sequence identity with equine, feline, human, porcine, and rhesus and 69-77% with bovine, cotton rat, mouse, and rat with 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. Zucker, K. et al. (1994) Lymphokine Cytokine Res. 13:191.
    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
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