Human leptin binding domain - Cat no. LBD-2 GENE ID 3953

Recombinant human leptin binding domain (hLBD), is one polypeptide chain containing 208 amino acids and an additional Ala at N-terminus acids. It consists of the cytokine binding domain of leptin receptor (amino acids 428-635 of human leptin receptor and having a molecular mass of ~ 24.5 kDa, It was purified by proprietary chromatographic techniques (see Sandowski et al. J. Biol. Chem. (2002) 277(48):46304-9.

E. coli

 

 

White lyophilized (freeze-dried) powder

The protein was lyophilized from a concentrated (0.2 to 0.5 mg/ml) solution of Tris-HCl buffer, pH 9.0 with 150 mM NaCl and 10-fold (w/w) excess of glycine. This formulation allows full solubilization by adding water and gentle mixing and fully preserves the binding capacity of hLBD.

It is recommended to reconstitute the lyophilized hLBD in sterile water adjusted to pH 8-9, not less than 100µg/ml, which can then be further diluted to other aqueous solutions, preferably in presence of carrier protein.

Lyophilized hLBD should be stored desiccated below -18C. Upon reconstitution at > 0.1 hLBD mg/ml and up to 40.5 mg/ml mM and filter sterilization hLBD can be stored at 4C for several months. Sterile solutions at 0.5 mg/ml or less are stable at +4C for several months.

Purity: Greater than 99.0% as determined by:
(a) Gel filtration analysis.
(b) Analysis by reducing and non-reducing SDS-PAGE gel.

The sequence of the first six N-terminal amino acids was determined and was found to be Ala-Ile-Asp-Val-Asn-Ile

The purified hLBD (24K) consists of > 95% monomers as determined by gel-filtration chromatography.

PLR’s hLBDL is fully biologically active as evidenced by high affinity binding of mammalian leptins at 1:1 molar ratio.

Less than 0.1 ng/µg (IEU/µg) of hLBD

Protein quantitation was carried out by UV spectroscopy at 280 nm using the absorbency value of 2.45 as the extinction coefficient for a 0.1% (1mg/ml) solution at pH 9.0. This value is calculated by the PC GENE computer analysis program of protein sequences (IntelliGenetics).

This material is offered by PLR for laboratory research