Certificate of Analysis and Data Sheet
Source:
E.Coli
Catalog No.
CYT-355
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 rat leptin, one polypeptide chain containing 146 amino and additional Ala at N-terminus acids and having a molecular mass of ~ 16 kDa, rLEP was mutated, resulting in L39A/D40A/F41A mutant that was purified by proprietary chromatographic techniques.
Physical Appearance:
Sterile Filtered White lyophilized (freeze-dried) powder.
Formulation:
The protein was lyophilized from a concentrated (0.85mg/ml) solution with 0.003mM NaHCO3.
Solubility:
It is recommended to reconstitute the lyophilized art LEP mutant in sterile water or sterile 0.4% NaHCO3 adjusted to pH 8-9, not less than 100ug/ml, which can then be further diluted with other aqueous solutions.
Stability:
Lyophilized rLEP mutant although stable at room temperature for several weeks, should be stored desiccated below -18
0C. Upon reconstitution at > 0.1 rat LEP mutant mg/ml and up to 2 mM and filter sterilization rLEP 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 99.0% as determined by:
(a) Gel filtration analysis.
(b) Analysis by reducing and non-reducing SDS-PAGE 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 rat LEP triple antagonist (16K) consists of > 99% monomers as determined by gel-filtration chromatography.
Biological Activity:
ProSpec旧 rLEP 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 rat LEP triple antagonist
Protein content:
Protein quantization was carried out by UV spectroscopy at 280 nm using the absorbency value of 0.21 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. Effect of smoking on neonatal and maternal serum and breast milk leptin levels.
Endocr Res 2005;31(3):177-83
2. Relationship between serum leptin levels and body composition and markers of malnutrition in nondiabetic patients on peritoneal dialysis or hemodialysis.
J Chin Med Assoc 2005 Dec;68(12):566-70
3. Relationship between insulin-like growth factors (IGF-I and IGF-II), IGF-binding proteins (IGFBP-3, IGFBP-2), leptin and anthropometric parameters (height, body mass index) during antileukaemic treatment in children.
Rocz Akad Med Bialymst 2005;50:208-11
4. Body composition, dehydroepiandrosterone sulfate and leptin concentrations in girls approaching menarche.
J Pediatr Endocrinol Metab 2005 Oct;18(10):975-83
5. Leptin levels in boys with pubertal gynecomastia.
J Pediatr Endocrinol Metab 2005 Oct;18(10):929-34
6. Plasma leptin concentrations in postmenopausal women with osteoporosis.
Endocr Res 2005;31(2):133-8