Canine TNF-alpha Biotinylated Antibody

  (1 citations)     
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  • 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
Product Datasheets

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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 Example
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.
    Reconstitution Buffer Available
  • 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 degreesC as supplied. 1 month, 2 to 8 degreesC under sterile conditions after reconstitution. 6 months, -20 to -70 degreesC 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
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.

1 Citations: Showing 1 - 1

  1. A Vaccine Therapy for Canine Visceral Leishmaniasis Promoted Significant Improvement of Clinical and Immune Status with Reduction in Parasite Burden
    Authors: BM Roatt, RD Aguiar-Soa, LE Reis, JM Cardoso, FA Mathias, RC de Brito, SM da Silva, NF Gontijo, SA Ferreira, JG Valenzuela, R Corrêa-Oli, RC Giunchetti, AB Reis
    Front Immunol, 2017;8(0):217.
    Species: Canine
    Sample Type: Cell Culture Supernates
    Application: ELISA detection

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Isotype Controls
Description Application Cat# Citations Images  

Normal Goat IgG Biotinylated Control

Ctrl BAF108 19
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Reconstitution Buffers
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Reconstitution Buffer 1 (PBS)

RB01
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Staining Reagents
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Streptavidin-Alkaline Phosphatase

ELISA(Det) AR001 4
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Streptavidin-Phycoerythrin

Flow F0040 7
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NorthernLights Streptavidin NL557

Flow , IHC , ICC NL999 4
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Streptavidin-Allophycocyanin

Flow F0050 1
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NorthernLights Streptavidin NL493

Flow , IHC , ICC NL997 2
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NorthernLights Streptavidin NL637

Flow , IHC , ICC NL998 2
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Avidin-Fluorescein

Flow F0030 1
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Supplemental Cell Selection Products
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MagCellect Streptavidin Ferrofluid

MAG999B 4
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MagCellect Streptavidin Ferrofluid

MAG999 4
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Supplemental ELISA Products
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Streptavidin-HRP

ELISA DY998 51  
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EvenCoat Streptavidin Coated Plates

ELISA CP004
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EvenCoat Streptavidin Coated Plates, 5 Pack

ELISA CP003 3
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