Detects recombinant cotton rat TNF-alpha in direct ELISAs and Western blots. In direct ELISAs, approximately 50% cross-reactivity with recombinant human (rh) EDA-A2 is observed, 5% cross-reactivity with rhTRAIL and recombinant mouse (rm) TRAIL is observed, and no cross‑reactivity with rhTNF-alpha, rmTNF-alpha, rhAPRIL, rhBAFF, rhEDA-A2, rhGITR Ligand, rhLIGHT, rhOX40 Ligand, rhTWEAK, rhVEGI, or rmTRANCE is observed.
Monoclonal Mouse IgG2B Clone # 159811
Protein A or G purified from hybridoma culture supernatant
E. coli-derived recombinant cotton rat TNF-alpha Leu1-Leu156 Accession # AAL18818
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.
Recombinant Cotton Rat TNF‑ alpha (Catalog # 1011-CR)
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 0.02-0.08 µg/mL in the presence of 1 ng/mL Recombinant Cotton 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 Cotton Rat TNF‑ alpha Antibody. Recombinant Cotton Rat TNF‑ alpha (Catalog # 1011-CR) 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 Cotton Rat TNF‑ alpha (0.5 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Cotton Rat TNF‑ alpha Monoclonal Antibody (Catalog # MAB1011). The ND50 is typically 0.05- 0.3 µg/mL in the presence of the metabolic inhibitor actinomycin D (1 µg/mL).
Preparation and Storage
Reconstitute at 0.5 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-α), 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-α is produced by a wide variety of immune and epithelial cell types. Human TNF-α consists of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 177 aa extracellular domain (ECD). Within the ECD, human TNF-α shares 97% aa sequence identity with rhesus and 71% - 92% with bovine, canine, cotton rat, equine, feline, mouse, porcine, and rat TNF-α. The 26 kDa type 2 transmembrane protein is assembled intracellularly to form a noncovalently linked homotrimer. Ligation of this complex induces reverse signaling that promotes lymphocyte costimulation but diminishes monocyte responsiveness.
Cleavage of membrane bound TNF-α by TACE/ADAM17 releases a 55 kDa soluble trimeric form of TNF-α. TNF-α trimers bind the ubiquitous TNF RI and the hematopoietic cell-restricted TNF RII, both of which are also expressed as homotrimers. TNF-α regulates lymphoid tissue development through control of apoptosis. It also promotes inflammatory responses by inducing the activation of vascular endothelial cells and macrophages. TNF-α is a key cytokine in the development of several inflammatory disorders. It contributes to the development of type 2 diabetes through its effects on insulin resistance and fatty acid metabolism.
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