Leptin tA Human
Leptin Antagonist Triple Mutant Human Recombinant
Cat. No.
BT22030
Source
Escherichia coli.
Synonyms
Appearance
White lyophilized (freeze-dried) powder.
Purity
Greater than 98.0% as determined by:
(a) Gel filtration analysis.
(b) Analysis by SDS-PAGE.
(a) Gel filtration analysis.
(b) Analysis by SDS-PAGE.
Usage
THE BioTek'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.
Shipped with Ice Packs 
In Stock 
Description
Leptin Antagonist Triple Mutant Human Recombinant is a single non-glycosilated polypeptide chain containing 146 amino and additional Ala at N-terminus acids and having a molecular weight of 16 kDa, Leptin was mutated, resulting in L39A/D40A/F41A. Leptin Antagonist Triple Mutant Human Recombinant was purified by proprietary chromatographic techniques.
Product Specs
Description
Leptin Antagonist Triple Mutant Human Recombinant is a single, non-glycosylated polypeptide chain consisting of 146 amino acids and an additional alanine residue at the N-terminus. With a molecular weight of 16 kDa, this variant features three mutations (L39A/D40A/F41A). Purification is achieved through proprietary chromatographic techniques.
Physical Appearance
White, lyophilized powder.
Formulation
The protein was lyophilized from a solution at a concentration of 1 mg/ml in 0.0045 mM sodium bicarbonate (NaHCO3).
Solubility
To reconstitute the lyophilized Leptin Antagonist Triple Mutant Human Recombinant, it is recommended to dissolve it in sterile 0.4% sodium bicarbonate (NaHCO3) at a concentration of at least 100 µg/ml. This solution can be further diluted in other aqueous solutions as needed.
Stability
Lyophilized Leptin Antagonist Triple Mutant Human Recombinant remains stable at room temperature for several weeks. However, for long-term storage, it should be kept desiccated below -18°C. Once reconstituted at a concentration between 0.1 mg/ml and 2 mg/ml and sterilized by filtration, the mutant protein can be stored at 4°C or even room temperature for several weeks, making it suitable for extended infusion studies utilizing osmotic pumps. At lower concentrations, adding a carrier protein like 0.1% HSA or BSA is advisable. Avoid repeated freeze-thaw cycles.
Purity
Purity exceeds 98.0% as determined by:
(a) Gel filtration analysis.
(b) SDS-PAGE analysis.
Biological Activity
THE BioTek's Leptin triple antagonist effectively inhibits the proliferation of BAF/3 cells stably transfected with the long form of the human leptin receptor, which is induced by leptin. Furthermore, it inhibits various leptin effects in multiple in vitro bioassays.
Protein Content
Protein quantification was performed using UV spectroscopy at a wavelength of 280 nm. An absorbency value of 0.88 was employed as the extinction coefficient for a 0.1% (1 mg/ml) solution at pH 8.0. This value was calculated utilizing the PC GENE computer analysis program for protein sequences (IntelliGenetics).
Source
Escherichia coli.
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.
Product Science Overview
Introduction
Leptin is a hormone predominantly made by adipose cells and enterocytes in the small intestine that helps to regulate energy balance by inhibiting hunger. It is often referred to as the “satiety hormone” or “fat hormone.” Leptin’s primary target is the hypothalamus in the brain where it regulates appetite and energy expenditure.
Leptin Antagonist Triple Mutant
The Leptin Antagonist Triple Mutant (Human Recombinant) is a modified form of the human leptin protein. This recombinant protein is engineered to act as an antagonist, meaning it blocks the action of natural leptin. The triple mutant refers to three specific amino acid substitutions in the leptin protein: L39A, D40A, and F41A .
Structural Characteristics
- Amino Acid Sequence: The Leptin Antagonist Triple Mutant is a single non-glycosylated polypeptide chain containing 146 amino acids, with an additional alanine (Ala) at the N-terminus .
- Molecular Weight: The molecular weight of this recombinant protein is approximately 16 kDa . However, due to its pegylation (attachment of polyethylene glycol), it exhibits an apparent molecular weight of 35.6 kDa in SDS-PAGE and over 200 kDa in gel-filtration .
- Pegylation: The protein is mono-pegylated with a 20 kDa PEG, which enhances its stability and solubility .
Production and Purification
Biological Activity
Applications
The Leptin Antagonist Triple Mutant is primarily used in research to study leptin signaling pathways and to investigate the physiological roles of leptin in energy balance, appetite regulation, and metabolic processes. It is also used to explore potential therapeutic applications for conditions related to leptin signaling, such as obesity and metabolic disorders.
Storage and Stability
- Lyophilized Form: The protein is lyophilized from a concentrated solution with NaHCO3 and is stable at room temperature for up to three weeks .
- Reconstitution: It is recommended to reconstitute the lyophilized protein in sterile 0.4% NaHCO3 adjusted to pH 8-9 .
- Storage Conditions: Upon reconstitution, the protein should be stored at 4°C for short-term use (2-7 days) and below -18°C for long-term storage. It is advisable to add a carrier protein (0.1% HSA or BSA) to prevent freeze-thaw cycles .
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