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Recombinant Mouse TNF RI/TNFRSF1A Fc Chimera Protein

R&D Systems, part of Bio-Techne | Catalog # 430-RI

R&D Systems, part of Bio-Techne
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Carrier Free
430-RI-050/CF

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With Carrier
430-RI-050

Key Product Details

Source

NS0

Accession #

Structure / Form

Disulfide-linked homodimer

Conjugate

Unconjugated

Applications

Bioactivity

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse TNF RI/TNFRSF1A protein
Mouse TNF RI
(Leu30-Ala212)
Accession # P25118
DIEGRMD Human IgG1
(Pro100-Lys330)
6-His tag
N-terminus C-terminus

Purity

>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Leu30

Predicted Molecular Mass

48.6 kDa (monomer)

SDS-PAGE

70 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 0.75-4.5 ng/mL in the presence of 0.1 ng/mL recombinant mouse TNF-alpha.

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: 430-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.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
Carrier Free: 430-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.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

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). 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 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

  1. Pfeffer, K. (2003) Cytokine Growth Factor Rev. 14:185.
  2. Hehlgans, T. and K. Pfeffer (2005) Immunology 115:1.
  3. Chan, F.K. et al. (2000) Science 288:2351.
  4. Lewis, M. et al. (1991) Proc. Natl. Acad. Sci. USA 88:2830.
  5. Peschon, J.J. et al. (1998) J. Immunol. 160:943.
  6. Banner, D.W et al. (1993) Cell 73: 431.
  7. Turner, M.D. et al. (2012) Biosci. Rep. 32:105.
  8. Legler, D.F. et al. (2003) Immunity 18:655.
  9. Tellier, E. et al. (2006) Exp. Cell Res. 312:3969.
  10. Xanthoulea, S. et al. (2004) J. Exp. Med. 200:367.
  11. Jin, L. et al. (2000) J. Immunol. 165:5153.
  12. Gomez, M.I. et al. (2006) J. Biol. Chem. 281:20190.
  13. Islam, A. et al. (2006) J. Biol. Chem. 281:6860.
  14. Garton, K.J. et al. (2006) J. Leukoc. Biol. 79:1105.
  15. McDermott, M.F. et al. (1999) Cell 97:133.

Long Name

Tumor Necrosis Factor Receptor I

Alternate Names

CD120a, TNFRI, TNFRSF1A

Entrez Gene IDs

7132 (Human); 21937 (Mouse); 25625 (Rat); 102135285 (Cynomolgus Monkey)

Gene Symbol

TNFRSF1A

UniProt

Additional TNF RI/TNFRSF1A Products

Product Documents for Recombinant Mouse TNF RI/TNFRSF1A Fc Chimera Protein

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant Mouse TNF RI/TNFRSF1A Fc Chimera Protein

For research use only

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