Description:
Recombinant chicken leptin binding domain (chLBD), 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 420-626 of chicken leptin receptor and having a molecular mass of ~ 24.5 kDa, It was purified by proprietary chromatographic techniques (see Niv-Spector et al. Biochemical Journal (2005) 390:475-484.
Source:
E. coli
Lot Number:
Quantity Shipped:
Physical Appearance:
White lyophilized (freeze-dried) powder or sterile 0.05% solution.
Formulation:
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 chLBD.
Solubility:
It is recommended to reconstitute the lyophilized chLBD in sterile water at pH 8-9, not less than 100µg/ml, which can then be further diluted to other aqueous solutions, preferably in presence of carrier protein.
Stability:
Lyophilized chLBD should be stored desiccated below -18C. Upon reconstitution at > 0.1 chLBD mg/ml and up to 40.5 mg/ml mM and filter sterilization chLBD 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.
(a) Gel filtration analysis.
(b) Analysis by reducing and non-reducing SDS-PAGE gel.
Amino Acid Sequence:
The sequence of the first six N-terminal amino acids was determined and was found to be Ala-Ile-Asp-Val-Asn-Ile
Dimers and Aggregates:
The purified chLBD (24K) consists of > 95% monomers as determined by gel-filtration chromatography.
Biological Activity:
PLR’s chLBDL is fully biologically active as evidenced by high affinity binding of mammalian leptins at 1:1 molar ratio.
Endotoxin:
Less than 0.1 ng/µg (IEU/µg) of chLBD
Protein content:
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).
Usage:
This material is offered by PLR for laboratory research
Date Shipped: