| Reactivity | MuSpecies 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 0.75-4.5 ng/mL in the presence of 0.1 ng/mL recombinant mouse TNF-alpha . |
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| Source | Mouse myeloma cell line, NS0-derived mouse TNF RI/TNFRSF1A protein
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| Accession # | |||||||||
| N-terminal Sequence | Leu30 |
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| Structure / Form | Disulfide-linked homodimer |
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| Protein/Peptide Type | Recombinant Proteins |
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| Gene | Tnfrsf1a |
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| Purity | >90%, by SDS-PAGE under reducing conditions and visualized by silver stain. |
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| Endotoxin Note | <0.10 EU per 1 μg of the protein by the LAL method. |
| Dilutions |
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| Theoretical MW | 48.6 kDa (monomer). 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. |
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| SDS-PAGE | 70 kDa, reducing conditions |
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| Publications |
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| Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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| Buffer | Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein. |
| Purity | >90%, by SDS-PAGE under reducing conditions and visualized by silver stain. |
| Reconstitution Instructions | Reconstitute at 100 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 and TNFRSF1A) is a 55 kDa type I transmembrane protein member of the TNF receptor superfamily, designated TNFRSF1A (1, 2). Mouse TNF RI is a 454 amino acid (aa) protein that contains a 21 aa signal sequence, a 191 aa extracellular domain (ECD) with a PLAD (pre-ligand assembly domain) that mediates constitutive dimer/trimer formation, followed by four CRD (cysteine-rich domains), a 23 aa transmembrane domain, and a 219 aa cytoplasmic sequence that contains a neutral sphingomyelinase activation domain and a death domain (3, 4). The ECD of mouse TNF RI shares 70%, 88%, 67%, 70% and 64% aa sequence identity with human, rat, canine, feline and porcine TNF RI, respectively. Both TNF RI and TNF RII (TNFRSF1B) are widely expressed and contain four TNF-alpha trimer-binding CRD in their ECD. However, TNF RI is thought to mediate most of the cellular effects of TNF-alpha (3). It is essential for proper development of lymph node germinal centers and Peyer’s patches, and for combating intracellular pathogens such as Listeria (1, 2, 5). TNF RI is also a receptor for TNF-beta /TNFSF1B (lymphotoxin-alpha ) (6). TNF RI is stored in the Golgi and translocates to the cell surface following pro-inflammatory stimuli (7). TNF-alpha stabilizes TNF RI and induces its sequestering in lipid rafts, where it activates NF kappa B and is cleaved by ADAM-17/TACE (8, 9, 16). Release of the 28-34 kDa TNF RI ECD also occurs constitutively and in response to products of pathogens such as LPS, CpG DNA or S. aureus protein A (1, 10-12). Full-length TNF RI may also be released in exosome-like vesicles (13). Release helps to resolve inflammatory reactions, since it down-regulates cell surface TNF RI and provides soluble TNF RI to bind TNF-alpha (10, 14, 15). Exclusion from lipid rafts causes endocytosis of TNF RI complexes and induces apoptosis (1). Mutations of human TNF R1 can result in inflammatory episodes known as TRAPS (TNFR-associated periodic syndrome) (7).
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