protein

Pegylated mouse super leptin antagonist (mutant D23/L39A/D40A/F41A)

MBS400080

0.05 mg

225 USD

Recombinant Protein

Pegylated mouse super leptin antagonist (mutant D23/L39A/D40A/F41A)

Pegylated super leptin antagonist

Mouse pegylated super leptin antagonist (mutant D23L/L39A/D40A/F41A)

Escherichia Coli

The sequence of the N-terminal amino acids of pegylated recombinant super mouse leptin antagonist could not be determined indicating that the PEG moiety is attached to the N-terminus.

The purity of pegylated recombinant super mouse leptin antagonist is greater than 95.0% as determined by:. (a) Gel filtration analysis. (b) Analysis by reducing and non-reducing SDS-PAGE gel.

White lyophilized (freeze-dried) powder.

Lyophilized pegylated recombinant super mouse leptin antagonistal though stable at room temperature for several weeks, should be stored desiccated below - 20 degree C. Upon reconstitution of pegylated recombinant super mouse leptin antagonist at > 0.1 mg/ml and up to 2 mg/ml and filter sterilization pegylated recombinant super mouse leptin antagonist mutant can be stored at 4 degree 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.

Solubility: It is recommended to reconstitute the lyophilized pegylated recombinant super mouse leptin antagonist in sterile water or sterile 0.4% NaHCO3 adjusted to pH 8-9, not less than 100ug/ml, which can then be further diluted with other aqueous solutions. Protein Content: Protein quantization of recombinant super mouse leptin antagonist was carried out by UV spectroscopy at 280 nm using the absorbency value of 0.200 as the extinction coefficient for a 0.1% (1mg/ml) protein solution at pH 8.0. This value is calculated by the PC GENE computer analysis program of protein sequences (IntelliGenetics). Dimers and Aggregates: The purified pegylated recombinant super mouse leptin antagonist consists of > 90% monomers as determined by gel-filtration chromatography.

Endotoxin: The endotoxin content of recombinant super mouse leptin antagonist is less than 0.01 ng/ug (0.1 EU/ug of protein) of PEG-SMLA. Formulation: The pegylated recombinant super mouse leptin antagonist was lyophilized from a concentrated (0.65mg/ml) solution with 0.003mM NaHCO3. Biological Activity: pegylated recombinant super mouse leptin antagonist is capable of inhibiting leptin-induced proliferation of BAF/3 cells stably transfected with the long form of human leptin receptor. Pegylated recombinant super mouse leptin antagonist in vitro activity is 6-8 fold lower than the non-pegylated pegylated recombinant super mouse leptin antagonist but in vivo it has profound weight gain effect (as compared to the non-pegylated antagonist), resulting mainly from increased food intake. Its in vivo activity compared to that of PEG-MLA is 9-27 fold higher.

Leptin and leptin muteins

Mono-pegylated mouse super leptin antagonist (with 20 kDa PEG) is one polypeptide chain containing 146 amino, an additional Ala at N-terminus and one molecule of PEG 20 kDa at its N-terminus, having an expected molecular mass of ~ 35.6 kDa as determined by MS. However due to enlarged hydrodymanic volume pegylated recombinant super mouse leptin antagonist runs on the SDS-PAGE as a 55 kDa protein and in gel-filtration on Superdex 200 as over 200 kDa protein. Pegylated recombinant super mouse leptin antagonist half-life in circulation after SC injection was over 20 hours. Super mouse leptin antagonist (SMLA) was initially mutated, resulting in D23L/L39A/D40A/F41A mutant that was purified by proprietary chromatographic techniques. Its pegylation is similar to that described in Elinav et al. Endocrinology 150:3083-91 (2009) for pegylated recombinant mouse leptin antagonist.

0.05 mg

225 USD