Recombinant Human TNF RI/TNFRSF1A Fc Chimera Protein
R&D Systems, part of Bio-Techne | Catalog # 372-RI
Key Product Details
Source
NS0
Accession #
Structure / Form
Disulfide-linked homodimer
Conjugate
Unconjugated
Applications
Bioactivity
Product Specifications
Source
Mouse myeloma cell line, NS0-derived human TNF RI/TNFRSF1A protein
| Human TNF RI (Leu30-Thr211) & (Asp41-Thr211) Accession # P19438 |
IEGRMD | Human IgG1 (Pro100-Lys330) |
6 His tag |
| N-terminus | C-terminus | ||
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<0.01 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Leu30 & Asp41
Predicted Molecular Mass
48 kDa (monomer)
SDS-PAGE
60-66 kDa, reducing conditions
Activity
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 0.4‑2 ng/mL in the presence of 0.25 ng/mL Recombinant Human TNF‑ alpha (Catalog # 210-TA).
The ED50 for this effect is 0.4‑2 ng/mL in the presence of 0.25 ng/mL Recombinant Human TNF‑ alpha (Catalog # 210-TA).
Formulation, Preparation and Storage
Carrier Free
What does CF mean?CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
What formulation is right for me?In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
Carrier: 372-RI
| Formulation | Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein. |
| Reconstitution | Reconstitute at 100 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin. |
| Shipping | The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Carrier Free: 372-RI/CF
| Formulation | Lyophilized from a 0.2 μm filtered solution in PBS. |
| Reconstitution | Reconstitute at 100 μg/mL in sterile PBS. |
| Shipping | The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Background: TNF RI/TNFRSF1A
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). Human TNF RI is a 455 amino acid (aa) protein that contains a 21 aa signal sequence and 190 aa 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 221 aa cytoplasmic sequence that contains a neutral sphingomyelinase activation domain and a death domain (3, 4). The ECD of human TNF RI shows 70%, 69%, 80%, 80%, and 73% aa identity with mouse, rat, canine, feline and porcine TNF RI, respectively; and it shows 23% aa identity with the ECD of TNF RII. 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 kappaB 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).
References
- Pfeffer, K. (2003) Cytokine Growth Factor Rev. 14:185.
- Hehlgans, T. and K. Pfeffer (2005) Immunology 115:1.
- Chan, F.K. et al. (2000) Science 288:2351.
- Schall, T.J. et al. (1990) Cell 61:361.
- Peschon, J.J. et al. (1998) J. Immunol. 160:943.
- Banner, D.W et al. (1993) Cell 73: 431.
- Turner, M.D. et al. (2012) Biosci. Rep. 32:105.
- Legler, D.F. et al. (2003) Immunity 18:655.
- Tellier, E. et al. (2006) Exp. Cell Res. 312:3969.
- Xanthoulea, S. et al. (2004) J. Exp. Med. 200:367.
- Jin, L. et al. (2000) J. Immunol. 165:5153.
- Gomez, M.I. et al. (2006) J. Biol. Chem. 281:20190.
- Islam, A. et al. (2006) J. Biol. Chem. 281:6860.
- Garton, K.J. et al. (2006) J. Leukoc. Biol. 79:1105.
- McDermott, M.F. et al. (1999) Cell 97:133.
Long Name
Tumor Necrosis Factor Receptor I
Alternate Names
CD120a, TNFRI, TNFRSF1A
Gene Symbol
TNFRSF1A
UniProt
Additional TNF RI/TNFRSF1A Products
Product Documents for Recombinant Human TNF RI/TNFRSF1A Fc Chimera Protein
Product Specific Notices for Recombinant Human TNF RI/TNFRSF1A Fc Chimera Protein
For research use only
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