The recombinant human TNF-alpha protein is expressed in E. coli, whose amino acid sequence is derived from human TNF-α soluble form (NP_000585.2), (Val 77-Leu 233) was expressed, with an initial Met at the N-terminus.
The recombinant human TNF-α consists of 158 amino acids and has a predicted molecular mass of 17.4 kDa which is also estimated by SDS-PAGE.
Human TNF-alpha protein measured in a cytotoxicity assay using L929 mouse fibrosarcoma cells in the presence of the metabolic inhibitor actinomycin D. The ED50 for this effect is typically 20-100 pg/ml, corresponding to a specific activity of 3 x 10E6-2 x 10E7 units/mg.
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