Equine TNF-alpha Antibody

Name: Equine TNF-alpha Antibody
Brand: R&D Systems
SKU: AF1814
Price: 159 USD
Availability: InStock

R&D Systems | Catalog # AF1814

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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

Equine

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 equine TNF-alpha
Leu78-Leu234
Accession # NP_001075288

Specificity

Detects equine TNF‑alpha in direct ELISAs and Western blots. In Western blots, approximately 35% cross-reactivity with recombinant mouse TNF-alpha and recombinant rat TNF-alpha is observed.

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 Equine TNF-alpha Antibody

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

Recombinant Equine TNF-a (Catalog # 1814-ET) induces cytotoxicity in the the L-929 mouse fibroblast cell line in a dose-dependent manner (orange line), as measured by Resazurin (Catalog # AR002). Cytotoxicity elicited by Recombinant Equine TNF-a (1 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Equine TNF-a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1814). The ND₅₀ is typically 0.01-0.03 µg/mL in the presence of the metabolic inhibitor actinomycin D (1 µg/mL).

TNF‑ alpha in Equine PBMCs.

TNF-a was detected in immersion fixed equine peripheral blood mononuclear cells (PBMCs) using Goat Anti-Equine TNF-a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1814) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # 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 Equine TNF-alpha Antibody

Application

Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed equine peripheral blood mononuclear cells (PBMCs)

Western Blot

0.1 µg/mL
Sample: Recombinant Equine TNF‑ alpha (Catalog # 1814-ET)

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 0.01-0.03 µg/mL in the presence of 1 ng/mL Recombinant Equine 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). Equine TNF-alpha consisits of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 178 aa extracellular domain (ECD) (3). Within the ECD, equine TNF-alpha shares 69%‑88% aa sequence identity with bovine, canine, cotton rat, feline, human, mouse, porcine, rat, and rhesus 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 costimulation 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.
Su, X.Z. et al. (1991) Gene 107:319.
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)