TNF alpha Recombinant Protein

Product Specific Information

The mature, soluble form of porcine TNFa was expressed in Pichia pastoris, KM71H. GenBank Accession # P23563, amino acids 77 - 232. For protein verification, the expressed protein was N-terminal sequenced and begins with the following 10 amino acids: LRSSSQTSDK. Purity is >95% by SDS-PAGE analyses.

Activity: The biological activity of recombinant porcine TNFa was measured by its cytolytic effect on porcine PK(15) cells in the presence of Actinomycin D. The ED50 (or 1 unit of activity) for this effect typically falls into the range of 7 - 18 pg/ml.

Use: For in vitro applications. A dose-response assay should be performed to determine the optimal concentration for specific experiments.

Reconstitution and Dilution: Store lyophilized material at 4°C. Reconstitute in 0.5 ml of distilled or deionized water. Dilute in a buffer-containing carrier protein, aliquot, and store at or below -70°C, avoiding multiple freeze-thaw cycles. PBS with 1% BSA or a neutral media containing <10% FCS are suitable buffers.

Target Information

TNF alpha is a multifunctional proinflammatory cytokine that belongs to the tumor necrosis factor (TNF) superfamily. This cytokine is mainly secreted by macrophage and bind to its receptors, TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. TNF alpha is involved in the regulation of immune cells, cell proliferation, differentiation, apoptosis, lipid metabolism, and coagulation. TNF alpha exists as a multimer of two, three, or five noncovalently linked units, but shows a single 17 kDa band following SDS-PAGE under non-reducing conditions. Knockout studies in mice also suggested the neuroprotective function of TNF alpha, and has been observed to causes tumor necrosis when injected into tumor-bearing mice. Other functions of TNF-alpha include its role in the immune response to bacterial, viral, parasitic and certain fungal infections, as well as its role in the necrosis of specific tumors. TNF alpha causes cytolysis or cytostasis of certain transformed cells, being synergistic with interferon-gamma in its cytotoxicity. This cytokine has been implicated in a variety of diseases, including autoimmune diseases, insulin resistance, and cancer.