Leptin Antagonist Quadruple Mutant Ovine Recombinant
Leptin Antagonist Quadruple 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 ~ 16 kDa, Leptin was mutated, resulting in L39A/D40A/F41A/I42A mutant.
Leptin Antagonist Quadruple Mutant Ovine Recombinant was purified by proprietary chromatographic techniques.
Accession
Amino acid sequence
The sequence of the first five N-terminal amino acids was determined and was found to be Ala-Val-Pro-Ile-Arg.
Biological Activity
leptin qA is capable of inhibiting leptin-induced proliferation of BAF/3 cells stably transfected with the long form of ovine leptin receptor. It is also inhibits various leptin effects in several in vitro bioassays.
Formulation
The protein was lyophilized from a concentrated (1mg/ml) solution with 0.0045mM NaHCO3.
Physical Appearance
White lyophilized (freeze-dried) powder.
Protein content
Protein quantitation was carried out by UV spectroscopy at 280 nm using the absorbency value of 0.21 as the extinction coefficient for a 0.1% (1mg/ml) solution at pH 8.0. This value is calculated by the PC GENE computer analysis program of protein sequences (IntelliGenetics).
Purity
Greater than 98.0% as determined by:
(a) Gel filtration analysis.
(b) Analysis by SDS-PAGE.
Solubility
It is recommended to reconstitute the lyophilized Leptin-Antagonist Quadruple Mutant Ovine Recombinant in sterile 0.4% NaHCO3adjusted to pH 8-9, not less than 100ug/ml, which can then be further diluted to other aqueous solutions.
Source
Stability
Lyophilized Leptin-Antagonist Quadruple Mutant Ovine Recombinant although stable at room temperature for several weeks, should be stored desiccated below -18oC. Upon reconstitution at > 0.1 LEP-qA mutant mg/ml and up to 2 mM and filter sterilization Leptin mutant can be stored at 4oC 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.