Protein Laboratories Rehovot Ltd.
MTS-tagged ovine leptin - Cat no. LEP-8 GENE ID 443534
Description:
Recombinant ovine MTS-leptin, one polypeptide chain containing 157 amino and (extra 10 amino acids of the membrane translocating sequence Val-Leu-Leu-Pro-Val-Leu-Leu-Ala-Ala-Pro) derived from Kaposi virus and additional Ala at N-terminus acids . The resulting molecular mass of is ~ 17.5 kDa, MTS-ovine leptin was purified by proprietary chromatographic techniques.
Source:
E. coli
Physical Appearance:
White lyophilized (freeze-dried) powder
Formulation:
The MTS-ovine recombinant leptin was lyophilized from a concentrated (1mg/ml) solution with 0.02% NaHCO3.
Solubility:
It is recommended to reconstitute the lyophilized MTS-ovine recombinant leptin in sterile 0.4% NaHCO3 adjusted, not less than 100µg/ml, which can then be further diluted to other aqueous solutions.
Stability:
Lyophilized MTS-ovine recombinant leptin although stable at room temperature for several weeks, should be stored desiccated below -18 C. Upon reconstitution of MTS-ovine recombinant leptin at > 0.1 hLEP mg/ml and up to 2 mg and filter sterilization hLEP can be stored at +4C.
Purity:
The purity of MYS-ovine recombinant leptin is greater than 98.0% as determined by:
(a) Gel filtration analysis.
(b) Analysis by reducing and non-reducing SDS-PAGE gel.
(c) Gel-filtration chromatography under non denaturing conditions.
(a) Gel filtration analysis.
(b) Analysis by reducing and non-reducing SDS-PAGE gel.
(c) Gel-filtration chromatography under non denaturing conditions.
Amino Acid Sequence:
The sequence of the first five N-terminal amino acids ovine recombinant leptin was determined and was found to be Ala-Val-Leu-Leu-Pro.
Dimers and Aggregates:
The purified MTS-ovine recombinant leptin (17.5 K) consists of > 96% monomers as determined by gel-filtration chromatography.
Biological Activity:
PLR’s MTS-ovine recombinant leptin is fully biologically active as evidenced by inducing proliferation of BAF/3 cells stably transfected with the long form of human leptin receptor. In vivo experiments has shown potency equal but not higher that non tagged ovine leptin (A. Gertler, unpublished data).
Endotoxin:
The endotoxin content of MTS-ovine recombinant leptin is less than 0.1 ng/µg (IEU/µg).
Protein content:
Protein quantitation of MTS-ovine recombinant leptin was carried out by UV spectroscopy at 280 nm using the absorbency value of 0.18 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).
Usage:
MTS-ovine recombinant leptinis offered by PLR for laboratory research.