Canine TNF-alpha Antibody
R&D Systems, part of Bio-Techne | Catalog # MAB1507
Key Product Details
Validated by
Biological Validation
Species Reactivity
Validated:
Canine
Cited:
Canine
Applications
Validated:
ELISA Capture (Matched Antibody Pair), Immunocytochemistry, Neutralization
Cited:
ELISA Capture, ELISA Development, ELISA Development (Capture)
Label
Unconjugated
Antibody Source
Monoclonal Mouse IgG1 Clone # 236812
Product Specifications
Immunogen
E. coli-derived recombinant canine TNF-alpha
Val77-Leu233
Accession # P51742
Val77-Leu233
Accession # P51742
Specificity
Detects canine TNF-alpha in direct ELISAs. In direct ELISAs, 25-100% cross-reactivity with recombinant human TNF‑ alpha, recombinant porcineTNF-alpha, and recombinant rhesus macaque TNF-alpha is observed.
Clonality
Monoclonal
Host
Mouse
Isotype
IgG1
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Scientific Data Images for Canine TNF-alpha Antibody
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).
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.Applications for Canine TNF-alpha Antibody
Application
Recommended Usage
Immunocytochemistry
8-25 µg/mL
Sample: Immersion fixed canine peripheral blood mononuclear cells treated with calcium ionomycin and PMA
Sample: Immersion fixed canine peripheral blood mononuclear cells treated with calcium ionomycin and PMA
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.
Canine TNF-alpha Sandwich Immunoassay
Please Note: Optimal dilutions of this antibody should be experimentally determined.
Formulation, Preparation, and Storage
Purification
Protein A or G purified from hybridoma culture supernatant
Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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.
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
Alternate Names
Cachetin, DIF, TNF, TNF-A, TNFA, TNFalpha, TNFG1F, TNFSF1A, TNFSF2
Entrez Gene IDs
Gene Symbol
TNF
UniProt
Additional TNF-alpha Products
Product Documents for Canine TNF-alpha Antibody
Product Specific Notices for Canine TNF-alpha Antibody
For research use only
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