| Reactivity | CaSpecies Glossary |
| Applications | Bioactivity |
| Details of Functionality | Measured by its ability to inhibit the TNF-alpha mediated cytotoxicity in the L‑929 mouse fibroblast cells in the presence of the metabolic inhibitor actinomycin D. Matthews, N. and M.L. Neale (1987) in Lymphokines and Interferons, A Practical Approach. Clemens, M.J. et al. (eds): IRL Press. 221. The ED50 for this effect is 0.04-0.2 µg/mL in the presence of 15 ng/mL of rcaTNF-alpha . |
| Source | E. coli-derived canine TNF RI/TNFRSF1A protein Ile22-Thr211, with an N-terminal Met |
| Accession # | |
| N-terminal Sequence | Met |
| Protein/Peptide Type | Recombinant Proteins |
| Purity | >95%, by SDS-PAGE under reducing conditions and visualized by silver stain |
| Endotoxin Note | <0.01 EU per 1 μg of the protein by the LAL method. |
| Dilutions |
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| Theoretical MW | 21.6 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. |
| Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
| Buffer | Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein. |
| Purity | >95%, by SDS-PAGE under reducing conditions and visualized by silver stain |
| Reconstitution Instructions | Reconstitute at 10 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin. |
TNF receptor 1 (TNF RI; also called TNF R-p55/p60, TNFRSF1A and CD120a) is a type I transmembrane protein that belongs to the TNF receptor superfamily (1, 2). TNF RI is widely expressed and is present on the cell surface as a trimer of 55 kDa subunits. It serves as a receptor for both TNF-alpha and TNF-beta /lymphotoxin. Each subunit contains four TNF-alpha trimer-binding cysteine-rich domains (CRD) in its extracellular domain (ECD) (1 - 6). TNF-alpha binding to TNF R1 induces the sequestration of TNF RI in lipid rafts, where it activates NF kappa B and is cleaved by ADAM-17/TACE (7, 8). Release of the 28 - 34 kDa TNF RI ECD occurs constitutively, and in response to products of pathogens such as LPS, CpG DNA or S. aureus protein A (1, 7 - 12). Full-length TNF RI may also be released in exosome-like vesicles (12). This release helps to resolve inflammatory reactions as it down-regulates cell surface TNF RI and provides soluble TNF RI to bind TNF-alpha (6, 13, 14). Exclusion from lipid rafts causes endocytosis of TNF RI complexes and induces apoptosis (7, 15). Although there is a second receptor for TNF-alpha (TNF R2), TNF RI is thought to mediate most of the cellular effects of TNF-alpha (3). TNF R1 is essential for proper development of lymph node germinal centers and Peyer’s patches, and for combating intracellular pathogens such as Listeria monocytogenes (1 - 3). Canine TNF RI is a 452 amino acid (aa) protein (16). Based on human, its ECD will contain a PLAD domain that mediates constitutive trimer formation (6). The PLAD domain will be followed by four CRDs, a transmembrane domain, and a cytoplasmic sequence that contains a neutral sphingomyelinase activation domain and a death domain. The ECD of canine TNF RI shows approximately 87%, 75%, 80%, 66% and 67% aa identity with feline, porcine, human, rat and mouse TNF RI, respectively.
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