Invitrogen™ Rat TNF-alpha Recombinant Protein

Recombinant Protein

The mature, soluble form of rat TNF-alpha was expressed in Pichia pastoris, KM71H. GenBank Accession # P16599, amino acids 80 - 235. For protein verification, the expressed protein was N-terminal sequenced and begins with the following 10 amino acids: EAEALRSSSQ. Purity is >95% by SDS-PAGE analyses. Activity: The biological activity of recombinant rat TNFa was measured by its cytolytic effect on mouse WEHI-13VAR cells in the presence of Actinomycin D. The ED50 (or 1 unit of activity) for this effect is typically <5 pg/mL. For in vitro applications a dose-response assay should be performed to determine the optimal concentration for specific experiments. For protein verification, the expressed protein was N-terminal sequenced and begins with the following 10 amino acids: EAEALSSSQ. The EAEA portion showed incomplete cleavage of the alpha-Factor signal by the Ste 13 protease. 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.