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Canine TNF-alpha Antibody

Catalog #: AF1507
1 Citation Datasheet
Catalog # Availability Size / Price Qty
AF1507
AF1507-SP
Cytotoxicity Induced by TNF‑ alpha and Neutralization by Canine TNF‑ alpha Antibody.
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Citations (1)
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Canine TNF-alpha Antibody Summary

Species Reactivity
Canine
Specificity
Detects canine TNF-alpha direct ELISAs and Western blots.
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 Trehalose. *Small pack size (SP) is supplied either lyophilized or 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 Canine TNF‑ alpha (Catalog # 1507-CT)
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 0.4-1.6 µg/mL in the presence of 100 ng/mL Recombinant Canine 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.

Scientific Data

Neutralization Cytotoxicity Induced by TNF‑ alpha and Neutralization by Canine TNF‑ alpha Antibody. View Larger

Cytotoxicity Induced by TNF‑ alpha and Neutralization by Canine TNF‑ alpha Antibody. Recombinant Canine TNF-a (1507-CT) 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 Canine TNF-a (100 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Canine TNF-a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1507). The ND50 is typically 0.4-1.6 µg/mL in the presence of the metabolic inhibitor actinomycin D (1 µg/mL).

Immunocytochemistry TNF-a antibody in Canine PBMCs by Immunocytochemistry (ICC). View Larger

TNF‑ alpha in Canine PBMCs. TNF-a was detected in immersion fixed canine peripheral blood mononuclear cells (PBMCs) stimulated with PMA and calcium ionomycin using Goat Anti-Canine TNF-a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1507) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) 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.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store 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 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). 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); 102139631 (Cynomolgus Monkey); 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; TNFG1F; 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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Citation for Canine TNF-alpha Antibody

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 Citation: Showing 1 - 1

  1. Biomimetic nanoparticles drive the mechanism understanding of shear-wave elasticity stiffness in triple negative breast cancers to predict clinical treatment
    Authors: D Zheng, J Zhou, L Qian, X Liu, C Chang, S Tang, H Zhang, S Zhou
    Bioactive materials, 2022-11-03;22(0):567-587.

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