Cytotoxicity Induced by TNF‑ alpha and Neutralization by Porcine TNF‑ alpha Antibody. Recombinant Porcine TNF‑ alpha (Catalog #|
690‑PT) induces cytotoxicity in the the PK‑15 porcine kidney epithelial cell line in a dose-dependent manner (orange line). Cytotoxicity elicited by Recombinant Porcine TNF‑ alpha (0.05 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Porcine TNF‑ alpha Monoclonal Antibody (Catalog # MAB6903). The ND50 is typically
0.005-0.02 µg/mL in the presence of the metabolic inhibitor actinomycin D (1 µg/mL).
|TNF‑ alpha in Porcine PBMCs. TNF‑ alpha was detected in immersion fixed porcine peripheral blood mononuclear cells treated with calcium ionomycin and PMA using Mouse Anti-Porcine TNF‑ alpha Monoclonal Antibody (Catalog # MAB6903) at 25 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 cytoplasm. 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). Porcine TNF-alpha consisits of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 176 aa extracellular domain (ECD) (3). Within the ECD, porcine TNF-alpha shares 69%‑86% aa sequence identity with bovine, canine, cotton rat, equine, feline, human, mouse, 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).
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