Canine TNF-alpha Antibody

Name: Canine TNF-alpha Antibody
Brand: R&D Systems
SKU: AF1507
Price: 159 USD
Availability: InStock

R&D Systems | Catalog # AF1507

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Calculators
Background
Product Documents
Specific Notices
Research Areas
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Key Product Details

Species Reactivity

Canine

Applications

Western Blot, Neutralization, Immunocytochemistry

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

View all TNF-alpha Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant canine TNF-alpha
Val77-Leu233
Accession # P51742

Specificity

Detects canine TNF-alpha direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

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 (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).

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.

Applications for Canine TNF-alpha Antibody

Application

Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed canine peripheral blood mononuclear cells

Western Blot

0.1 µg/mL
Sample: Recombinant Canine TNF‑ alpha (Catalog # 1507-CT)

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.

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

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

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

7124 (Human); 21926 (Mouse); 24835 (Rat); 397086 (Porcine); 280943 (Bovine); 403922 (Canine); 102139631 (Cynomolgus Monkey); 100033834 (Equine); 493755 (Feline); 100009088 (Rabbit)