Catalog Number: AT-0001H-1000/CF
Human tumor necrosis factor alpha (TNF-α), also known as cachectin and TNFSF2, is the prototypic ligand of the TNF superfamily. TNF-α is produced by a wide variety of immune, epithelial, endothelial, and tumor cells1,2. It is a primary mediator of numerous immunologic functions, including hemorrhagic tumor necrosis/cytotoxicity, inflammation and regulation of antiviral and immune proliferative and activation responses. As a central player in the cytokine network, TNF-α has been implicated in a variety of disease states, including cachexia, endotoxic (septic) shock, acute respiratory distress syndrome and a number of necrotic, proliferative and autoimmune diseases. Human TNF-α encoded by a 7124 gene, is a type II integral membrane protein consisiting of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 177 aa extracellular domain (ECD). Within the ECD, human TNF-α shares 97% aa sequence identity with rhesus and 71%-92% with bovine, canine, cotton rat, equine, feline, mouse, porcine, and rat TNF-α. TNF-α is assembled intracellularly to form a noncovalently linked homotrimer which is expressed on the cell surface3,4. Shedding of membrane bound TNF-α by TACE/ADAM17 releases the bioactive cytokine, a 60 kDa soluble trimer of the TNF-α extracellular domain5-7. TNF-α binds the ubiquitous 55-60 kDa TNFR I8,9 and the hematopoietic cell-restricted 80 kDa TNFR II, both of which are also expressed as homotrimers1,2,10. The TNF-α transmembrane protein is proteolytically cleaved to yield a soluble protein, which subsequently forms a non-covalently linked homotrimer in solution11. The individual subunits of this homotrimer have a relative molecular mass each of 20 KDa. TNF-α binds two receptors TNFR I and TNFR II inducing NF-κB and MAPK signaling pathways. Recombinant human TNF-α is intended for use in cell culture applications.
Recombinant human TNF-α is produced in Chinese hamster ovary (CHO) cells transfected with the ECD sequence of the human TNF-α gene with an 8-His tag at C-terminals.
Source: Mammalian; Chinese hamster ovary (CHO) cells
Species: Human; Gene ID: 7124; UniProt ID: P01375
Molecular weight: 20.6 kDa including tags
Tags: 8-His tag at C-terminals
Concentration: determined by A280 with the Extinction coefficient of 1.044 l/g∙ cm-1 to produce 1 mg/vial, 200 μg/vial product form.
Amino acids: encoding the extracellular (57-233) domain of human TNF with an 8-His tag at C-terminals.
Amino Acid Sequence:
Formulation: Carrier free recombinant human TNF-α was lyophilized from a 0.22 mm filtered solution in sterile ammonium bicarbonate buffer solution (pH 7.4) (Catalog # AT-0001H/CF; carrier free product formulation) or Recombinant human TNF-α was lyophilized from a 0.22 mm filtered solution in sterile ammonium bicarbonate buffer solution (pH 7.4) and 5% trehalose containing (Catalog # AT-0001H; Stabilier product formulation).
– Purity greater than 98% as determined by SDS-PAGE.
– Homotrimer conformation as determined by SEC chromatography.
– TNF-α ELISA for TNF-α folding validation. -Biological activity is tested using L929 cells for cytotoxicity assay. -Endotoxin levels <0.05 EU/μg as determined using a kinetic chromogenic LAL assay.
This protocol uses Cell Proliferation Assay to determine bioactivity of TNF-α.
Recombinant human TNF-α in lyophilized carrier free powder
form is shipped at room temperature.
– It should be stored at -20 °C; stored at -80oC upon receiving for
long term storage.
– Preparation of stock solution (100 μg/ml):
1. Add 10 ml endotoxin-free water to 1000 μg of
recombinant human TNF-α.
2. Mix by pipetting. Do not vortex.
3. Prepare aliquots of recombinant human TNF-α. Avoid
4. Further dilutions can be prepared in the appropriate
– Note: Avoid repeated freeze-thaw cycles.
1.Zelova H, Hosek J. 2013. Inflamm Res 62: 641-51.
2.Juhasz K, Buzas K, Duda E. 2013. Expert Rev Clin Immunol 9: 335-48.
3.Tang P, Hung MC, Klostergaard J. 1996. Biochemistry 35: 8216-25.
4.Perez C, Albert I, DeFay K, Zachariades N, Gooding L, Kriegler M. 1990. Cell 63: 251-8.
5.Black RA, Rauch CT, Kozlosky CJ, Peschon JJ, Slack JL, et al. 1997. Nature 385: 729-33.
6.Moss ML, Jin SL, Milla ME, Bickett DM, Burkhart W, et al. 1997. Nature 385: 733-6.
7.Gearing AJ, Beckett P, Christodoulou M, Churchill M, Clements J, et al. 1994. Nature 370: 555-7.
8.Schall TJ, Lewis M, Koller KJ, Lee A, Rice GC, et al. 1990. Cell 61: 361-70.
9.Loetscher H, Pan YC, Lahm HW, Gentz R, Brockhaus M, et al. 1990. Cell 61: 351-9.
10.Loetscher H, Gentz R, Zulauf M, Lustig A, Tabuchi H, et al. 1991. J Biol Chem 266: 18324-9.
11.Kriegler M, Perez C, DeFay K, Albert I, Lu SD. 1988. Cell 53: 45-53.
|Human TNF-α ELISA Set||AT-hTNFα-ELISA001|
|Recombinant Human TNF‑α||AT-0001H-200/CF|
|Recombinant Human TNF‑α||AT-0001H-50|
|Recombinant Human TNF‑α||AT-0001H-200|
|Recombinant Human TNF‑α||AT-0001H-1000|
ATBM manufactures products for a number of intended uses. Please refer to the product label for the intended use statements for specific products. ATBM products contain chemicals which may be harmful if misused. Due care should be exercised with all Promega products to prevent direct human contact.
Each ATBM product is shipped with documentation stating specifications and other technical information. ATBM products are warranted to meet or exceed the stated specifications. ATBM’s sole obligation and the customer’s sole remedy is limited to replacement of products free of charge in the event products fail to perform as warranted. ATBM makes no other warranty of any kind whatsoever, and SPECIFICALLY DISCLAIMS AND EXCLUDES ALL OTHER WARRANTIES OF ANY KIND OR NATURE WHATSOEVER, DIRECTLY OR INDIRECTLY, EXPRESS OR IMPLIED, INCLUDING, WITHOUT LIMITATION, AS TO THE SUITABILITY, PRODUCTIVITY, DURABILITY, FITNESS FOR A PARTICULAR PURPOSE OR USE, MERCHANTABILITY, CONDITION, OR ANY OTHER MATTER WITH RESPECT TO ATBM PRODUCTS. In no event shall ATBM be liable for claims for any other damages, whether direct, incidental, foreseeable, consequential, or special (including but not limited to loss of use, revenue or profit), whether based upon warranty, contract, tort (including negligence) or strict liability arising in connection with the sale or the failure of ATBM products to perform in accordance with the stated specifications.