OLeptine tA PEG Recombinant eiwit

Source : Escherichia coli. Pegylated Leptin Antagonist Triple Mutant Ovine Recombinant is a single non-glycosilated polypeptide chain containing 146 amino and additional Ala at N-terminus acids and having a molecular mass of ~ 35.6 kDa, Leptin was mutated, resulting in L39A/D40A/F41A mutant. However due to enlarged hydrodymanic volume it runs on the SGS-Page as 48 kDa protein and in gel-filtration on Superdex 200 as over 200kDa protein. Leptin Antagonist Triple Mutant Ovine Recombinant Mono-Pegylated with 20kDa PEG and was purified by proprietary chromatographic techniques.

Greater than 95.0% as determined by: (a) Gel filtration analysis. (b) Analysis by SDS-PAGE.

The protein was lyophilized from a concentrated (0.65mg/mL) solution with 0.003mM NaHCO3.

Lyophilized Leptin Antagonist Triple Mutant Ovine Recombinant although stable at room temperature for several weeks, should be stored desiccated below -18°C. Upon reconstitution at > 0.1 Lep-tA mutant mg/ml and up to 2mM and filter sterilization Leptin mutant can be stored at 4°C or even room temperature for several weeks making it suitable for long term infusion studies using osmotic pumps. At lower concentration addition of a carrier protein (0.1% HSA or BSA) is suggested. Please prevent freeze-thaw cycles.

It is recommended to reconstitute the lyophilized Leptin-Antagonist Triple Mutant Ovine Recombinant in sterile water or sterile 0.4% NaHCO3 adjusted to pH 8, not less than 100μg/ml, which can then be further diluted to other aqueous solutions. Pegylated triple antagonist is capable of inhibiting leptin-induced proliferation of BAF/3 cells stably transfected with the long form of human leptin receptor. Its in vitro activity is 6-8 fold lower than the non-pegylated antagonist but in vivo it has profound weight gain effect (as compared to the non-pegylated antagonist like in mouse leptin antagonists), resulting mainly from increased food intake.