Certificate of Analysis and Data Sheet

Source:
E.Coli

Catalog No.
CYT-239

Background:

Leptin inhibits food intake and stimulates energy expenditure. Leptin also has thermogenic actions and regulates enzymes of fatty acid oxidation. Severe hereditary obesity in rodents and humans is caused by defects in leptin production .In addition to its critical role in the physiologic regulation of body weight leptin has a variety of other physiologic and pathologic functions resembling those of cytokines . These functions include the regulation of hematopoiesis , angiogenesis , wound healing , inflammation , and immune responses.

Description :

Recombinant Ovine Leptin produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 146 amino acids and having a molecular mass of 16,130 Dalton.
Recombinant Ovine Leptin is purified by proprietary chromatographic techniques.

Physical Appearance:

Sterile Filtered White lyophilized (freeze-dried) powder.

Formulation:

Recombinant Ovine Leptin was lyophilized from a concentrated (1mg/ml) solution with 0.0045mM NaHCO3.

Solubility:

It is recommended to reconstitute the lyophilized Leptin in sterile 18M-cm H2O not less than 100ug/ml, which can then be further diluted to other aqueous solutions.

Stability:

Lyophilized Leptin although stable at room temperature for 3 weeks, should be stored desiccated below -18 C. Upon reconstitution rOLeptin should be stored at 4 C between 2-7 days and for future use below -18 C. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).

Please prevent freeze-thaw cycles.

Purity:

Greater than 95.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Anion-exchange FPLC.
(c) Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel.

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.

Dimers and aggregates:

Less than 1% as determined by silver-stained SDS-PAGE gel analysis.

Biological Activity:

ProSpec's Leptin is fully biologically active when compared to standards. The ED50, calculated by the leptin-dependant stimulation of Human OB-R transfected murine BaF3 indicator cells is 0.33-0.05 ng/ml.

Endotoxin:

Less than 0.1 ng/ug (IEU/ug) of Leptin.

Protein content:

Protein quantitation was carried out by two independent methods:
1. UV spectroscopy at 280 nm using the absorbency value of 0.2 as the extinction coefficient for a 0.1% (1mg/ml) solution. This value is calculated by the PC GENE computer analysis program of protein sequences (IntelliGenetics).
2. Analysis by RP-HPLC,using calibrated solution of Ovine Leptin as a Reference Standard.

Usage:

Prospec's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.

Gene:

Name:LEP
Synonyms:OB

Protein synonyms/aliases:

Leptin (Obesity factor).

Protein Family:

Belongs to the leptin family.

Protein Domains:

Domains:
•IPR000065   Obesity factor

Related proteins:

•Recombinant Human Leptin
•Recombinant Murine Leptin
•Recombinant Rat Leptin

Protein links:

Precursor- Protein structure and amino acid sequence:

Latest Publications:

1. Identification of the hydrophobic strand in the A-B loop of leptin as major binding site III: implications for large-scale preparation of potent recombinant human and ovine leptin antagonists.
Biochem J 2005 Oct 15;391(Pt 2):221-30
2. Chronic administration of recombinant ovine leptin in growing beef heifers: effects on secretion of LH, metabolic hormones, and timing of puberty.
J Anim Sci 2004 Oct;82(10):2930-6
3. Overexpression of ovine leptin in Pichia pastoris: physiological yeast response to leptin production and characterization of the recombinant hormone.
Yeast 2004 Feb;21(3):249-63
4. The late gestation increase in circulating ACTH and cortisol in the fetal sheep is suppressed by intracerebroventricular infusion of recombinant ovine leptin.
J Endocrinol 2002 Aug;174(2):259-66
5. Central infusion of recombinant ovine leptin normalizes plasma insulin and stimulates a novel hyper secretion of luteinizing hormone after short-term fasting in mature beef cows.
Biol Reprod 2002 May;66(5):1555-61
6. Effect of intravenous infusion of recombinant ovine leptin on feed intake and serum concentrations of GH, LH, insulin, IGF-1, cortisol, and thyroxine in growing prepubertal ewe lambs.
Domest Anim Endocrinol 2002 Apr;22(2):103-12

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中華民國95年06月06日更新