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. The specific activity of recombinant human TNF-alpha GMP is >4.3 x 107 IU/mg, which is calibrated against the human TNF-alpha WHO International Standard (NIBSC code: 12/154).
Recombinant Human TNF-alpha GMP Protein, CF Scientific Data Examples
Recombinant Human TNF-alpha GMP Protein Bioactivity
GMP-grade Recombinant Human TNF-alpha (Catalog # 210-GMP) induces cytotoxicity in the L-929 mouse fibroblast cell line in the presence of the metabolic inhibitor actinomycin D. The ED50 is 25‑100 pg/mL.
Recombinant Human TNF-alpha GMP Protein SDS-PAGE
1 μg/lane of GMP-grade Recombinant Human TNF-alpha (Catalog # 210-GMP) was resolved by SDS-PAGE with silver staining, under reducing (R) conditions, showing a single band at 17 kDa.
Recombinant Human TNF-alpha GMP Protein Mass Spectrometry
MALDI-TOF analysis of GMP-grade Recombinant Human TNF-alpha (Catalog # 210‑GMP). The major peak at 17348 corresponds to the calculated molecular mass without an N-terminal Met, 17353 Da. The minor peak at 17480 corresponds to the calculated molecular mass with an N-terminal Met, 17484 Da. The peak at 17565 is a matrix-associated artifact of the MALDI-TOF.
Formulation, Preparation and Storage
What does CF mean?
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our
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
Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitute at 0.1-1 mg/mL in PBS.
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.
A minimum of 6 months when stored at ≤ -20 °C as supplied. Refer to lot specific COA for the Use by Date.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
3 months, ≤ -20 °C under sterile conditions after reconstitution.
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).
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Moss, M.L. et al. (1997) Nature 385:733.
Gearing, A.J.H. et al. (1994) Nature 370:555.
Schall, T.J. et al. (1990) Cell 61:361.
Loetscher, H. et al. (1990) Cell 61:351.
Dembic, Z. et al. (1990) Cytokine 2:231.
Smith, C.A. et al. (1990) Science 248:1019.
Loetscher, H. et al. (1991) J. Biol. Chem. 266:18324.
Pinckard, J.K. et al. (1997) J. Biol. Chem. 272:10784.
Engelmann, H. et al. (1990) J. Biol. Chem. 265:1531.
Which Brands are Currently Available on bio-techne.com? R&D Systems and Tocris Bioscience products are available to purchase through bio-techne.com. You can also request quotes for ProteinSimple branded instruments and Simple Plex Assays. ProteinSimple branded instrument consumables and ACD branded products will be available on bio-techne.com in the near future. Novus Biologicals branded products are not currently available on bio-techne.com and can be found at novusbio.com.
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