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Rat pegylated leptin antagonist (mutant L39A/D40A/F41A) - Cat no. PELAN-2 GENE ID 25608

Description:
Mono-pegylated (with 20 kDa PEG) recombinant rat leptin antagonist 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 rat leptin antagonist runs on the SGS-Page as 48 kDa protein and on gel-filtration Superdex 200 column as over 100 kDa protein. The half-life of recombinant rat leptin antagonist in circulation after SC injection was over 20 hours. Recombinant rat leptin was initially mutated, resulting in L39A/D40A/F41A mutant that was purified by proprietary chromatographic techniques according to Salomon et al (2006) Protein Expression and Purification 47, 128–136.
Source:
Escherichia coli.
Physical Appearance:
White lyophilized (freeze-dried) powder.
Formulation:
The recombinant pegylated rat leptin antagonist was lyophilized from a concentrated (0.65mg/ml) solution with 0.003mM NaHCO3.
Solubility:
It is recommended to reconstitute the pegylated rat leptin antagonist in sterile water or sterile 0.4% NaHCO3 adjusted to pH 8-9, not less than 100µg/ml, which can then be further diluted with other aqueous solutions.
Stability:
Lyophilized pegylated rat leptin antagonist although stable at room temperature for several weeks, should be stored desiccated below -18C. Upon reconstitution of pegylated rat leptin antagonist at > 0.1 mg/ml and up to 2 mg/ml and filter sterilization pegylated rat leptin antagonist can be stored at 4C or even room temperature for several weeks making it suitable for long term infusion studies using osmotic pumps. At lower concentration of pegylated rat leptin antagonist addition of a carrier protein (0.1% HSA or BSA) is suggested. Please prevent freeze-thaw cycles.
Purity:
The purity of pegylated rat leptin antagonist is greater than 99.0% as determined by:
(a) Gel filtration analysis.
(b) Analysis by reducing and non-reducing SDS-PAGE gel.
Amino Acid Sequence:
The sequence of N-terminal amino acids of pegylated rat leptin antagonist could not be determined indicating that the PEG is attached to the N-terminus.
Dimers and Aggregates:
The purified pegylated rat leptin antagonist (16K) consists of > 99% monomers as determined by gel-filtration chromatography.
Biological Activity:
PLR’s pegylated rat leptin antagonist is capable of inhibiting leptin-induced proliferation of BAF/3 cells stably transfected with the long form of human leptin receptor. The in vitro activity of pegylated rat leptin antagonist is 5-6 fold lower than the non-pegylated antagonist but in vivo it has profound weight gain effect (as compared to the non-pegylated antagonist), resulting mainly from increased food intake.
Endotoxin:
Less than 0.05 ng/µg (EU/µg) of pegylated rat leptin antagonist .
Protein content:
Protein quantitation of pegylated rat leptin antagonist was carried out by UV spectroscopy at 280 nm using the absorbency value of 0.201 as the extinction coefficient for a 0.1% (1mg/ml) solution of protein at pH 8.0. This value is calculated by the PC GENE computer analysis program of protein sequences (IntelliGenetics).
Usage:
PLR's pegylated rat leptin antagonist is furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals. It is for Research Purposes Only – Commercial Use has been Licensed Exclusively to a Third Party.