Certificate of Analysis & Data Sheet

Recombinant rat leptin, one polypeptide chain containing 146 amino and additional Ala at N-terminus acids and having a molecular mass of ~ 16 kDa. Recombinant rat leptin was mutated, resulting in L39A/D40A/F41A mutant that was purified by proprietary chromatographic techniques, according to Salomon et al (2006) Protein Expression and PuriWcation 47, 128–136.

Escherichia coli.

White lyophilized (freeze-dried) powder.

Recombinant rat leptin was lyophilized from a concentrated (0.85mg/ml) solution with 0.003mM NaHCO3.

It is recommended to reconstitute the lyophilized recombinant 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.

Lyophilized recombinant rat leptin antagonist although stable at room temperature for several weeks, should be stored desiccated below -18C. Upon reconstitution of recombinant rat leptin antagonist at > 0.1 rLEP mutant mg/ml and up to 2 mg/ml and filter sterilization recombinant 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 recombinant rat leptin antagonist addition of a carrier protein (0.1% HSA or BSA) is suggested. Please prevent freeze-thaw cycles.

The purity of recombinant rat leptin antagonist is greater than 98.0% as determined by:
(a) Gel filtration analysis.
(b) Analysis by reducing and non-reducing SDS-PAGE gel.

The sequence of the first five N-terminal amino acids of recombinant rat leptin antagonist was determined and was found to be Ala-Val-Pro-Ile-Gln

The purified recombinant rat leptin antagonist (16K) consists of > 97% monomers as determined by gel-filtration chromatography.

PLR’s recombinant rat leptin antagonist is capable of inhibiting leptin-induced proliferation of BAF/3 cells stably transfected with the long form of human leptin receptor. It also inhibits various leptin effects in several in vitro bioassays.

Less than 0.1 ng/µg (IEU/µg) of recombinant rat leptin antagonist.

Protein quantitation of recombinant 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 at pH 8.0. This value is calculated by the PC GENE computer analysis program of protein sequences (IntelliGenetics).

PLR's recombinant rat leptin antagonist is furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives, local application or household chemicals. It is for Research Purposes Only – Commercial Use has been Licensed Exclusively to a Third Party.