Certificate of Analysis and Data Sheet
Source:
E.Coli
Catalog No.
CYT-352
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, one polypeptide chain containing 146 amino and additional Ala at N-terminus acids and having a molecular mass of ~ 16 kDa, hLEP was mutated, resulting in L39A/D40A/F41A mutant was purified by proprietary chromatographic techniques.
Physical Appearance:
Sterile Filtered White lyophilized (freeze-dried) powder.
Formulation:
The protein was lyophilized from a concentrated (1mg/ml) solution with 0.0045mM NaHCO3.
Solubility:
It is recommended to reconstitute the lyophilized hLEP mutant in sterile 0.4% NaHCO3 adjusted to pH 8-9, not less than 100ug/ml, which can then be further diluted to other aqueous solutions.
Stability:
Lyophilized hLEP mutant although stable at room temperature for several weeks, should be stored desiccated below -18
0C. Upon reconstitution at > 0.1 hLEP mutant mg/ml and up to 2 mM and filter sterilization hLEP mutant can be stored at 4
0C or even room temperature for several weeks making it suitable for long term infusion studies using osmotic pumps. At lower concentration addition of a carrier protein (0.1% HSA or BSA) is suggested.
Please avoid freeze-thaw cycles.
Purity:
Greater than 98.0% as determined by:
(a) Gel filtration analysis.
(b) 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-Gln
Dimers and aggregates:
The purified hLEP triple antagonist (16K) consists of > 95% monomers as determined by gel-filtration chromatography.
Biological Activity:
ProSpec旧 humanLEP triple 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.
Endotoxin:
Less than 0.1 ng/ug (IEU/ug) of human LEP triple antagonist
Protein content:
Protein quantitation was carried out by UV spectroscopy at 280 nm using the absorbency value of 0.88 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).
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.
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 infl ammatory marker levels in humans with leptin sufficiency or excess.
J Clin Endocrinol Metab 2005 Mar;90(3):1618-24