Recombinant Human TNF-alpha (HEK293-expressed) Protein, CF Best Seller

R&D Systems, part of Bio-Techne | Catalog # 10291-TA

Analyzed by SEC-MALS
R&D Systems, part of Bio-Techne
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Key Product Details

Source

HEK293

Accession #

P01375

Conjugate

Unconjugated

Applications

Bioactivity

View all TNF-alpha Proteins and Enzymes »

Product Specifications

Source

Human embryonic kidney cell, HEK293-derived human TNF-alpha protein
Val77-Leu233

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.

N-terminal Sequence Analysis

Val77

Predicted Molecular Mass

17 kDa

SDS-PAGE

14-18 kDa, under reducing conditions

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 20-100 pg/mL.

Reviewed Applications

Read 3 reviews rated 4.3 using 10291-TA in the following applications:

Scientific Data Images for Recombinant Human TNF-alpha (HEK293-expressed) Protein, CF

Recombinant Human TNF-alpha (HEK293-expressed) Protein SEC-MALS

Recombinant human TNF-alpha (Catalog # 10291-TA) has a molecular weight (MW) of 52.3 kDa as analyzed by SEC-MALS, suggesting that this protein is a homotrimer.  MW may differ from predicted MW due to post-translational modifications (PTMs) present (i.e. Glycosylation).
Recombinant Human TNF-alpha (HEK293-expressed) Protein Bioactivity

Recombinant Human TNF-alpha (HEK293-expressed) Protein Bioactivity

Recombinant Human TNF‑ alpha (HEK293-expressed) (Catalog # 10291-TA) induces cytotoxicity in the L‑929 mouse fibroblast cell line in the presence of the metabolic inhibitor actinomycin D. The ED50 for this effect is 20‑100 pg/mL.
Recombinant Human TNF-alpha (HEK293-expressed) Protein SDS-PAGE

Recombinant Human TNF-alpha (HEK293-expressed) Protein SDS-PAGE

2 μg/lane of Recombinant Human TNF-alpha (HEK293-expressed) (Catalog # 10291-TA) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 14-18 kDa.

Formulation, Preparation and Storage

10291-TA
Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution Reconstitute at 500 μg/mL in 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-alpha

Tumor necrosis factor alpha (TNF-alpha), also known as cachectin and TNFSF2, 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

Alternate Names

Cachetin, DIF, TNF, TNF-A, TNFA, TNFalpha, TNFG1F, TNFSF1A, TNFSF2

Entrez Gene IDs

7124 (Human); 21926 (Mouse); 24835 (Rat); 397086 (Porcine); 280943 (Bovine); 403922 (Canine); 102139631 (Cynomolgus Monkey); 100033834 (Equine); 493755 (Feline); 100009088 (Rabbit)

Gene Symbol

TNF

UniProt

P01375 (Human)

Additional TNF-alpha Products

Product Documents for Recombinant Human TNF-alpha (HEK293-expressed) Protein, CF

Product Datasheets

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Product Specific Notices for Recombinant Human TNF-alpha (HEK293-expressed) Protein, CF

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