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H8916

Sigma-Aldrich

Tumor Necrosis Factor-α human

TNF-α, recombinant, expressed in HEK 293 cells, suitable for cell culture

Synonym(s):
TNF-α
CAS Number:
MDL number:
NACRES:
NA.77

Quality Level

biological source

human

recombinant

expressed in HEK 293 cells

assay

≥95% (SDS-PAGE)

form

lyophilized powder

potency

≤1.0 ng/mL ED50

quality

endotoxin tested

mol wt

17 kDa (glycosylated)
~17.4 kDa

packaging

pkg of 10 μg

storage condition

avoid repeated freeze/thaw cycles

application(s)

cell culture | mammalian: suitable

impurities

≤1 EU/μg

UniProt accession no.

storage temp.

−20°C

Gene Information

human ... TNF(7124)

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General description

The TNFα (tumor necrosis factor α) gene is mapped to human chromosome 6p21.33. TNFα is a member of TNF superfamily. TNF-α has a palmitoyl group in the cysteine residue and is phosphorylated in the transmembrane region serine residue.

Application

Tumor Necrosis Factor-a human has been used:
  • as a permeability inducing agent for endothelial cell monolayer permeability assay
  • as a reactive oxygen species inducer in primary rat cardiac microvascular endothelial cells (RCMVECs)
  • in the activation of nuclear factor kappa B (NF-κB) in human embryonic kidney cells (HEK293), neuroblastoma SH-SY5Y cells and HeLa cells
  • in the stimulation of the human keratinocyte cell line(HaCaT) and human coronary artery endothelial cells (HCAECs)
Tumor Necrosis Factor-α (TNF-α) human has been used
  • In induction of netting neutrophils by anti-neutrophil cytoplasmic antibody and to study its effect on platelet activation and formation of monomeric C-reactive protein.
  • To study the effect of TNF-α on miR-221 and fractalkine expression.
  • To induce inflammatory cell responses.
  • In NF-κB luciferase reporter assay.

Packaging

10 μg in poly bottle

Biochem/physiol Actions

Tumor Necrosis Factor-α (TNF-α) is a potent pro-inflammatory cytokine that plays a role in the rheumatoid arthritis pathology, psoriatic arthritis (PsA) and psoriasis. It stimulates interleukins IL-1 and IL-6. Polymorphism in the TNF-α gene is associated with destructive arthropathy in PsA. The post-translational modifications in TNF-a is crucial for its functionality.
Tumore necrosis factor-α (TNF-α), also known as cachectin, is expressed as a 26 kDa membrane bound protein and is then cleaved by TNF-α converting enzyme (TACE) to release the soluble 17 kDa monomer, which forms homotrimers in circulation. TNF-α plays roles in antitumor activity, immune modulation, inflammation, anorexia, cachexia, septic shock, viral replication and hematopoiesis. TNF-α is expressed by a great variety of cells, with numerous inductive and suppressive agents. Primarily, TNF-α is produced by macrophages in response to immunological challenges such as bacteria (lipopolysaccharides), viruses, parasites, mitogens and other cytokines. TNF-α is cytotoxic for many transformed cells (its namesake activity) but in normal diploid cells, it can stimulate proliferation (fibroblasts), differentiation (myeloid cells) or activation (neutrophils). TNF-α also shows antiviral effects against both DNA and RNA viruses and it induces production of several other cytokines. Although TNF-α is used in clinical trials as an antitumor agent, Sigma′s cytokine, growth factor and hormone products are for research only. TNF-α and the related molecule TNF-β (LT-α) share close structural homology with 28% amino acid sequence identity and both activate the same TNF receptors, TNFR1 and TNFR2. Mouse and human TNF-α share 79% amino acid sequence identity. Unlike human TNF-α, the mouse form is N-glycosylated.

Preparation Note

Tumor Necrosis Factor-α (TNF-α) is expressed in human HEK 293 cells and has been shown to be predominantly a glycosylated, non-covalently linked homotrimer with a molecular mass of 51 kDa (gel filtration). Production in human 293 cells offers authentic glycosylation. Glycosylation contributes to stability in cell growth media and other applications.

Analysis Note

The specific activity was determined by the dose-dependent cytotoxity of the TNF alpha sensitive cell line L-929 in the presence of actinomycin D.

Storage Class Code

11 - Combustible Solids

WGK

WGK 2

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis

Certificate of Origin

  1. Which document(s) contains shelf-life or expiration date information for a given product?

    If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.

  2. How do I get lot-specific information or a Certificate of Analysis?

    The lot specific COA document can be found by entering the lot number above under the "Documents" section.

  3. How do I find price and availability?

    There are several ways to find pricing and availability for our products. Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers.

  4. What is the Department of Transportation shipping information for this product?

    Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product. 

  5. How should product H8916, Tumor Necrosis Factor-alpha human be dissolved?

    Product H8916 is soluble in sterile PBS or sterile water with 0.1 % endotoxin free human serum albumin. H8916 can be made without addition of albumin providing the protein concentration is high, 200-500 μg/ml. Since the vial contains 10 μg, then a volume of either sterile PBS or sterile water in the range of 20 -50 μl will be added to give the 200 -500 μg/ml protein concentration.

  6. What is the solution stablity of H8916, Tumor Necrosis Factor-alpha human?

    A stock solution of H8916 can be kept at a concentration of 200-500 ug/ml (micrograms/ml) for up to 1 week at 4 °C.  For long term storage, aliquot into single use size and store at -80 °C. Please avoid freeze thaw cycles as this can effect the activity.

  7. How should product H8916, Tumor Necrosis Factor-alpha human be dissolved?

    Product H8916 is soluble in sterile PBS or sterile water with 0.1 % endotoxin free human serum albumin. H8916 can be made without addition of albumin providing the protein concentration is high, 200-500 μg/ml. Since the vial contains 10 μg, then a volume of either sterile PBS or sterile water in the range of 20 -50 μl will be added to give the 200 -500 μg/ml protein concentration.

  8. What is the difference between H8916 and T6674, Tumor Necrosis Factor-α human?

    Both products are recombinant Tumor Necrosis Factor-α human, however, they are expressed in different cell types.  T6674 is expressed in Escherichia coli and H8916 is expressed in human HEK 293 cells. When recombinant human proteins are produced in human cells they are more authentic in terms of proper folding, phosphorylation, and glycosylation of the protein.  Human proteins expressed in  Escherichia coli do not undergo these proper post-translational modifications.

  9. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

C-reactive protein enhances activation of coagulation system and inflammatory response through dissociating into monomeric form in antineutrophil cytoplasmic antibody-associated vasculitis
Xu PC, et al.
BMC Immunology, 16.1 (2015)
Takahiko Horiuchi et al.
Rheumatology (Oxford, England), 49(7), 1215-1228 (2010-03-03)
Transmembrane TNF-alpha, a precursor of the soluble form of TNF-alpha, is expressed on activated macrophages and lymphocytes as well as other cell types. After processing by TNF-alpha-converting enzyme (TACE), the soluble form of TNF-alpha is cleaved from transmembrane TNF-alpha and...
Monitoring Cellular Stress Responses using Integrated High-Frequency Impedance Spectroscopy and Time-Resolved ELISA
Charwat V, et al.
Analyst, 139.20, 5271-5282 (2014)
LSHGD: A database for human leprosy susceptible genes
Doss CGP, et al.
Genomics, 100, 162-166 (2012)
Association between TNFβ - 308 G/A polymorphism and oral lichen planus (OLP): a meta-analysis
Zhou Y and Vieira AR
Journal of Applied Oral Science : Revista FOB, 26 (2018)

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