Detects rat TNF‑ alpha in direct ELISAs and Western blots. In Western blots, approximately 25% cross-reactivity with recombinant porcine TNF-alpha is observed and less than 1% cross-reactivity with recombinant human (rh) TNF-beta, rhTNF sRI, and rhTNF sRII is observed. In direct ELISAs, approximately 40% cross-reactivity with recombinant canine TNF-alpha, recombinant feline TNF-alpha, and recombinant rhesus macaque TNF-alpha is observed.
Neutralizes the biological activity of rat TNF-alpha. This antibody will also neutralize the biological activity of murine TNF-alpha, but weak neutralization of rhTNF-alpha and no neutralization of rhTNF-beta was observed.
Polyclonal Goat IgG
E. coli-derived recombinant rat TNF-alpha Leu80-Leu235 Accession # P16599
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
<0.10 EU per 1 μg of the antibody by the LAL method.
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 0.15-0.75 µg/mL in the presence of 0.25 ng/mL Recombinant Rat 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.
Cytotoxicity Induced by TNF‑ alpha and Neutralization by Rat TNF‑ alpha Antibody. Recombinant Rat TNF‑ alpha (Catalog # 510-RT) induces cytotoxicity in the the L‑929 mouse fibroblast cell line in a dose-dependent manner (orange line), as measured by Resazurin. Cytotoxicity elicited by Recombinant Rat TNF‑ alpha (0.25 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Rat TNF‑ alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-510-NA). The ND50 is typically 0.15‑0.75 µg/mL in the presence of the metabolic inhibitor actinomycin D (1 µg/mL).
TNF‑ alpha in Rat Splenocytes. TNF‑ alpha was detected in immersion fixed rat splenocytes using Goat Anti-Rat TNF‑ alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-510-NA) 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 cytoplasmic. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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.
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). Rat TNF-alpha consisits of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 179 aa extracellular domain (ECD) (3). Within the ECD, rat TNF-alpha shares 94% aa sequence identity with mouse and 69% - 76% with bovine, canine, cotton rat, equine, feline, human, porcine, 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 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).
Idriss, H.T. and J.H. Naismith (2000) Microsc. Res. Tech. 50:184.
Hehlgans, T. and K. Pfeffer (2005) Immunology 115:1.
Estler, H.C. et al. (1992) Biol. Chem. Hoppe-Seyler 373:271.
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.
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The data collected includes not only links to publications in PubMed,
but also provides information about sample types, species, and experimental conditions.
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