Recombinant Human TNF-alpha, Biotinylated Protein

Catalog # Availability Size / Price Qty
BT210-010
Recombinant Human TNF-alpha, Biotinylated Protein Bioactivity
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Citations (5)
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Recombinant Human TNF-alpha, Biotinylated Protein Summary

Product Specifications

Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<0.10 EU per 1 μg of the protein by the LAL method.
Activity
Measured in a cytotoxicity assay using 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 25-100 pg/mL.
Source
E. coli-derived human TNF-alpha protein
Val77-Leu233, with and without an N-terminal Met
Accession #
N-terminal Sequence
Analysis
Val77
Structure / Form
Homotrimer. Biotinylated protein via amines
Predicted Molecular Mass
17 kDa (unlabeled)
SDS-PAGE
17 kDa, reducing conditions

Product Datasheets

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BT210 (with carrier)

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BT210/CF (carrier free)

Discontinued Product

BT210

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.
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.

Scientific Data

Bioactivity Recombinant Human TNF-alpha, Biotinylated Protein Bioactivity View Larger

Both Biotinylated Recombinant Human TNF-a (Catalog # BT210) and unlabeled Recombinant Human TNF-a (Catalog # 210-TA) promotes cytotoxicity in L-929 mouse fibroblast cells in the presence of the metabolic inhibitor actinomycin D. The ED50 for this effect is 25-100 pg/mL. The similarity in activity highlights that the biotinylated protein is fully functional.

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Background: TNF-alpha

Tumor necrosis factor alpha (TNF-alpha ), also known as cachectin and TNFSF1A, is the prototypic ligand of the TNF superfamily. It is a pleiotropic molecule that plays a central role in inflammation, immune system development, apoptosis, and lipid metabolism (1, 2). Human TNF-alpha consisits of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 177 aa extracellular domain (ECD) (3). Within the ECD, human TNF-alpha shares 97% aa sequence identity with rhesus and 71%-92% with bovine, canine, cotton rat, equine, feline, mouse, porcine, and rat TNF-alpha. TNF-alpha is produced by a wide variety of immune, epithelial, endothelial, and tumor cells (1, 2). TNF-alpha is assembled intracellularly to form a noncovalently linked homotrimer which is expressed on the cell surface (4). Cell surface TNF-alpha can induce the lysis of neighboring tumor cells and virus infected cells, and it can generate its own downstream cell signaling following ligation by soluble TNFR I (2, 5). Shedding of membrane bound TNF-alpha by TACE/ADAM17 releases the bioactive cytokine, a 55 kDa soluble trimer of the TNF-alpha extracellular domain (6-8). TNF-alpha binds the ubiquitous 55-60 kDa TNF RI (9, 10) and the hematopoietic cell-restricted 80 kDa TNF RII (11, 12), both of which are also expressed as homotrimers (1, 2, 13). Both type I and type II receptors bind TNF-alpha with comparable affinity (14), although only TNF RI contains a cytoplasmic death domain which triggers the activation of apoptosis. Soluble forms of both types of receptors are released and can neutralize the biological activity of TNF-alpha (15).

References
  1. Zelova, H. and J. Hosek (2013) Inflamm. Res. 62:641.
  2. Juhasz, K. et al. (2013) Expert Rev. Clin. Immunol. 9:335.
  3. Pennica, D. et al. (1984) Nature 312:724.
  4. Tang, P. et al. (1996) Biochemistry 35:8216.
  5. Perez, C. et al. (1990) Cell 63:251.
  6. Black, R.A. et al. (1997) Nature 385:729.
  7. Moss, M.L. et al. (1997) Nature 385:733.
  8. Gearing, A.J.H. et al. (1994) Nature 370:555.
  9. Schall, T.J. et al. (1990) Cell 61:361.
  10. Loetscher, H. et al. (1990) Cell 61:351.
  11. Dembic, Z. et al. (1990) Cytokine 2:231.
  12. Smith, C.A. et al. (1990) Science 248:1019.
  13. Loetscher, H. et al. (1991) J. Biol. Chem. 266:18324.
  14. Pinckard, J.K. et al. (1997) J. Biol. Chem. 272:10784.
  15. Engelmann, H. et al. (1990) J. Biol. Chem. 265:1531.
Long Name
Tumor Necrosis Factor alpha
Entrez Gene IDs
7124 (Human); 21926 (Mouse); 24835 (Rat); 397086 (Porcine); 280943 (Bovine); 403922 (Canine); 102139631 (Cynomolgus Monkey); 100033834 (Equine); 493755 (Feline); 100009088 (Rabbit)
Alternate Names
APC1 protein; Cachectin; Cachetin; DIF; TNF; TNF, monocyte-derived; TNFA; TNF-A; TNFalpha; TNF-alpha; TNF-alphacachectin; TNFATNF, macrophage-derived; TNFG1F; TNFSF1A; TNFSF2; TNFSF2TNF superfamily, member 2; tumor necrosis factor (TNF superfamily, member 2); tumor necrosis factor alpha; Tumor necrosis factor ligand superfamily member 2; tumor necrosis factor; tumor necrosis factor-alpha

Citations for Recombinant Human TNF-alpha, Biotinylated Protein

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

5 Citations: Showing 1 - 5
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  1. Biodistribution and function of coupled polymer-DNA origami nanostructures
    Authors: Joseph, N;Shapiro, A;Gillis, E;Barkey, S;Abu-Horowitz, A;Bachelet, I;Mizrahi, B;
    Scientific reports
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  2. Roles for ADAM17 in TNF-R1 Mediated Cell Death and Survival in Human U937 and Jurkat Cells
    Authors: J Fritsch, J Frankenhei, L Marischen, T Vadasz, A Troeger, S Rose-John, D Schmidt-Ar, W Schneider-
    Cells, 2021-11-10;10(11):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  3. Regulation of CYLD activity and specificity by phosphorylation and ubiquitin-binding CAP-Gly domains
    Authors: PR Elliott, D Leske, J Wagstaff, L Schlicher, G Berridge, S Maslen, F Timmermann, B Ma, R Fischer, SMV Freund, D Komander, M Gyrd-Hanse
    Cell Reports, 2021-10-05;37(1):109777.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  4. Soft extracellular matrix enhances inflammatory activation of mesenchymal stromal cells to induce monocyte production and trafficking
    Authors: SW Wong, S Lenzini, MH Cooper, DJ Mooney, JW Shin
    Sci Adv, 2020-04-08;6(15):eaaw0158.
    Species: Human
    Sample Types: Whole Cells
    Applications: Cell Culture
  5. ST6Gal-I sialyltransferase promotes tumor necrosis factor (TNF)-mediated cancer cell survival via sialylation of the TNF receptor 1 (TNFR1) death receptor
    Authors: AT Holdbrooks, CM Britain, SL Bellis
    J. Biol. Chem., 2017-12-12;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay

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