Certificate of Analysis and Data Sheet

Source:
E.Coli

Catalog No.
CYT-228

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 Human Leptin produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 147 amino acids and having a molecular mass of 16,240 Dalton.
Recombinant Leptin is purified by proprietary chromatographic techniques.

Physical Appearance:

Sterile Filtered White lyophilized (freeze-dried) powder.

Formulation:

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

Solubility:

The lyophilized Leptin is very soluble in water and most aqueous buffers below and above the isoelectric point.

Stability:

Lyophilized Leptin although stable at room temperature, should be stored desiccated below 0 C. Reconstituted rHuLeptin is best stored refrigerated at 4 C.
Please avoid 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 Met-Val-Pro-Ile-Gln.

Dimers and aggregates:

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

Biological Activity:

ProSpec's Recombinant Human 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 0.5-1.6 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 0.878 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 a calibrated solution of 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 precursor (Obesity factor)
    (Obese protein).

Protein Family:

Belongs to the leptin family.

Protein Domains:

Domains:
•IPR000065   Obesity factor

Related proteins:

•Recombinant Murine Leptin
•Recombinant Rat Leptin
•Recombinant Ovine Leptin

Protein links:

Precursor- Protein structure and amino acid sequence:

Latest Publications:

1. Human leptin: an adipocyte hormone with weight-regulatory and endocrine functions.
Semin Vasc Med 2005 Feb;5(1):15-24
2. The long-term effect of recombinant methionyl human leptin therapy on hyperandrogenism and menstrual function in female and pituitary function in male and female hypoleptinemic lipodystrophic patients.
Metabolism 2005 Feb;54(2):255-63
3. Association analysis of the Gln223Arg polymorphism in the human leptin receptor gene, and traits related to obesity in Mexican adolescents.
J Hum Hypertens 2005 May;19(5):341-6
4. Effect of administration of recombinant human leptin during the neonatal period on the plasma concentration and gene expression of leptin in the piglet.
Biol Neonate 2005;87(1):1-7
5. Recombinant methionyl human leptin administration activates signal transducer and activator of transcription 3 signaling in peripheral blood mononuclear cells in vivo and regulates soluble tumor necrosis factor-alpha receptor levels in humans with relative leptin deficiency.
J Clin Endocrinol Metab 2005 Mar;90(3):1625-31
6. Recombinant methionyl human leptin administration to achieve high physiologic or pharmacologic leptin levels does not alter circulating i nflammatory marker levels in humans with leptin sufficiency or excess.
J Clin Endocrinol Metab 2005 Mar;90(3):1618-24

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中華民國95年06月06日更新