Leptin Human, N82K
Greater than 98.0% as determined by:
(a) Gel filtration analysis.
(b) Analysis by SDS-PAGE.
Description
Leptin N82K Human Recombinant produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 146 amino acids (and additional Ala at N-terminus acids) and having a molecular mass of 16kDa.
The Leptin N82K is purified by proprietary chromatographic techniques.
Product Specs
Leptin is a hormone that plays a crucial role in regulating energy balance and body weight. It achieves this by binding to the leptin receptor (LEPR), which activates several signaling pathways. In the hypothalamus, a brain region involved in appetite control, leptin acts to suppress appetite and increase energy expenditure. It also influences bone mass and the release of hormones from the hypothalamus and pituitary gland. In other parts of the body, leptin can increase metabolism, regulate the function of pancreatic beta cells (responsible for insulin production), and modulate immune responses.
Leptin N82K Human Recombinant, produced in E. coli, is a single-chain polypeptide that is not glycosylated (meaning it lacks attached sugar molecules). It consists of 146 amino acids, with an additional alanine (Ala) at the beginning of the chain (N-terminus). This protein has a molecular weight of 16 kDa.
The purification of Leptin N82K is achieved using specialized chromatographic methods.
The product is lyophilized (freeze-dried) from a concentrated solution (1mg/ml) containing 0.0045mM NaHCO3 (sodium bicarbonate).
To reconstitute the lyophilized Leptin N82K, it is recommended to dissolve it in sterile water or a 0.4% NaHCO3 solution adjusted to a pH of 8-9. The initial concentration should be at least 100 µg/ml, and this solution can be further diluted as needed in other aqueous solutions.
Lyophilized Leptin N82K remains stable at room temperature for up to 3 weeks. However, for long-term storage, it is recommended to store it desiccated (dry) at a temperature below -18°C. After reconstitution, the Leptin N82K solution should be stored at 4°C for a maximum of 2-7 days. For extended storage, adding a carrier protein like 0.1% HSA (human serum albumin) or BSA (bovine serum albumin) is advised. To maintain stability, avoid repeated freezing and thawing cycles.
The purity of Leptin N82K is greater than 98.0%, as determined by the following methods:
(a) Gel filtration analysis (a technique that separates molecules based on their size).
(b) SDS-PAGE analysis (a method that separates proteins based on their size and charge).
The biological activity of this product is less than 0.1%, as measured by its ability to stimulate the proliferation of BAF/3 cells that have been genetically modified to express the long form of the human leptin receptor.
Protein concentration was determined using UV spectroscopy at a wavelength of 280 nm. An extinction coefficient of 0.87 was used for a 0.1% (1mg/ml) solution at pH 8.0. This extinction coefficient was calculated using the PC GENE computer program (IntelliGenetics), which analyzes protein sequences.
The sequence of the first five N-terminal amino acids was determined and was found to be Ala-Val-Pro-Ile-Gln.
Product Science Overview
Leptin (Human Recombinant, N82K) is a mutant form of human leptin. It is produced in Escherichia coli (E. coli) and is a single, non-glycosylated polypeptide chain containing 146 amino acids, with an additional alanine at the N-terminus . The molecular mass of this recombinant leptin is approximately 16 kDa .
Leptin plays a crucial role in regulating energy balance and body weight control. After entering the circulation, leptin binds to the leptin receptor (LEPR), which results in the activation of several major signaling pathways . In the hypothalamus, leptin acts as an appetite-regulating factor that induces a decrease in food intake and an increase in energy consumption . It also regulates bone mass and the secretion of hypothalamo-pituitary-adrenal hormones . In the periphery, leptin increases basal metabolism, regulates pancreatic beta-cell function and insulin secretion, and affects innate and adaptive immunity .
Leptin (Human Recombinant, N82K) is typically provided as a sterile filtered, white lyophilized (freeze-dried) powder . It is recommended to reconstitute the lyophilized leptin in sterile water or 0.4% NaHCO3 adjusted to pH 8-9 . The lyophilized form is stable at room temperature for up to three weeks but should be stored desiccated below -18°C for long-term storage . Upon reconstitution, it should be stored at 4°C for short-term use (2-7 days) and below -18°C for long-term use . To prevent freeze-thaw cycles, it is recommended to add a carrier protein such as 0.1% HSA or BSA .
