Recombinant Human TNF-alpha Hyperactive Mutant Protein

R&D Systems, part of Bio-Techne | Catalog # 8599-TA

Mutant form with approximately 5-fold greater activity
R&D Systems, part of Bio-Techne
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8599-TA-010/CF
8599-TA-025/CF
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8599-TA-100/CF
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8599-TA-010
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Key Product Details

Source

E. coli

Accession #

P01375

Structure / Form

Non-covalent trimer

Conjugate

Unconjugated

Applications

Bioactivity

View all TNF-alpha Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human TNF-alpha protein
Lys87-Leu233 (Leu233Phe), with an N-terminal Met-Arg-Lys-Arg

Purity

>95%, by SDS-PAGE with silver staining.

Endotoxin Level

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

N-terminal Sequence Analysis

Met

Predicted Molecular Mass

17 kDa

SDS-PAGE

16 kDa, 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 2.5-12.5 pg/mL.

Scientific Data Images for Recombinant Human TNF-alpha Hyperactive Mutant Protein

Recombinant Human TNF-alpha Hyperactive Mutant Protein Bioactivity

Recombinant Human TNF-alpha Hyperactive Mutant Protein Bioactivity

Recombinant Human TNF-alpha Hyperactive Mutant (Catalog # 8599-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 2.5-12.5 pg/mL. This is an approximate 10-fold activity increase over the wild-type 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: 8599-TA
Formulation Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
Reconstitution Reconstitute at 100 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.
Carrier Free: 8599-TA/CF
Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution Reconstitute at 100 μg/mL in PBS.
Shipping The product is shipped with polar packs. 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 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). This recombinant protein contains an N-terminal insertion and a Leu233Phe substitution which have been shown to increase the bioactivity of TNF-alpha in vitro and in vivo (16).

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.
  16. Yan, Z. et al. (2006) Cytotherapy 8:415.

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

Additional TNF-alpha Products

Product Documents for Recombinant Human TNF-alpha Hyperactive Mutant Protein

Product Datasheets

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human TNF-alpha Hyperactive Mutant Protein

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