|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.
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).
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