Canine TNF-alpha Antibody

Catalog #: MAB1507
3 Citations Datasheet

Catalog #	Availability	Size / Price	Qty
MAB1507
MAB1507-SP

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

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All TNF-alpha Antibodies

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Citations (3)

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Canine TNF-alpha Antibody Summary

Species Reactivity

Canine

Specificity

Detects canine TNF-alpha in direct ELISAs. In direct ELISAs, 25-100% cross-reactivity with recombinant human TNF-alpha, recombinant porcine TNF-alpha, and recombinant rhesus macaque TNF-alpha is observed.

Source

Monoclonal Mouse IgG₁ Clone # 236812

Purification

Protein A or G purified from hybridoma culture supernatant

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

Immunocytochemistry

8-25 µg/mL

See below

Canine TNF-alpha Sandwich Immunoassay

Recommended Concentration

Reagent

ELISA Capture (Matched Antibody Pair)

2-8 µg/mL

Use in combination with:

Detection Reagent: Canine TNF‑ alpha Biotinylated Antibody (Catalog # BAF1507)

Standard: Recombinant Canine TNF-alpha Protein (Catalog # 1507-CT)

Neutralization

Measured by its ability to neutralize TNF‑ alpha -induced cytotoxicity in the L‑929 mouse fibroblast cell line. The Neutralization Dose (ND₅₀) is typically 1.25-5.0 µg/mL in the presence of 10 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

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Cytotoxicity Induced by TNF‑ alpha and Neutralization by Canine TNF‑ alpha Antibody. Recombinant Canine TNF-a (Catalog # 1507-CT) induces cytotoxicity in the the L-929 mouse fibroblast cell line in a dose-dependent manner (orange line). Cytotoxicity elicited by Recombinant Canine TNF-a (10 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Canine TNF-a Monoclonal Antibody (Catalog # MAB1507). The ND₅₀ is typically 1.25-5.0 µg/mL in the presence of the metabolic inhibitor actinomycin D (1 µg/mL).

Immunocytochemistry

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TNF‑ alpha in Canine PBMCs. TNF-a was detected in immersion fixed canine peripheral blood mononuclear cells (PBMCs) untreated or treated with calcium ionomycin and PMA using Mouse Anti-Canine TNF-a Monoclonal Antibody (Catalog # MAB1507) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cell secretion. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

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, 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

Idriss, H.T. and J.H. Naismith (2000) Microsc. Res. Tech. 50:184.
Hehlgans, T. and K. Pfeffer (2005) Immunology 115:1.
Zucker, K. et al. (1994) Lymphokine Cytokine Res. 13:191.
Tang, P. et al. (1996) Biochemistry 35:8216.
Eissner G. et al. (2004) Cytokine Growth Factor Rev. 15:353.
Black, R.A. et al. (1997) Nature 385:729.
Moss, M.L. et al. (1997) Nature 385:733.
Loetscher, H. et al. (1991) J. Biol. Chem. 266:18324.
Clark, I.A. (2007) Cytokine Growth Factor Rev. 18:335.
Romanatto, T. et al. (2007) Peptides 28:1050.
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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Related Research Areas

Citations 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.

3 Citations: Showing 1 - 3
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Phase I and II Clinical Trial Comparing the LBSap, Leishmuneï¿½, and Leish-Tecï¿½ Vaccines against Canine Visceral Leishmaniasis
Authors: RDO Aguiar-Soa, BM Roatt, FAS Mathias, LES Reis, JMO Cardoso, RCF Brito, HG Ker, R Corrêa-Oli, RC Giunchetti, AB Reis
Vaccines (Basel), 2020-11-17;8(4):.
Species: Canine
Sample Types: Cell Culture Supernates
Applications: ELISA Capture
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, 2017-03-07;8(0):217.
Species: Canine
Sample Types: Cell Culture Supernates
Applications: ELISA Development (Capture)
Expression of Toll-like receptors 2 and 9 in cells of dog jejunum and colon naturally infected with Leishmania infantum.
Authors: Figueiredo M, Amorim I, Pinto A, Barbosa V, Pinheiro L, Deoti B, Faria A, Tafuri W
BMC Immunol, 2013-05-14;14(0):22.
Species: Canine
Sample Types: Tissue Homogenates
Applications: ELISA Development