Leptin tA Rat, PEG

Leptin Antagonist Triple Mutant Pegylated Rat Recombinant
Cat. No.
BT22757
Source
Escherichia coli.
Synonyms
Appearance
White lyophilized (freeze-dried) powder.
Purity
Greater than 99.0% as determined by:
(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.
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Description

Leptin Antagonist Triple Mutant Rat Recombinant is a single non-glycosilated polypeptide chain containing 146 amino and additional Ala at N-terminus acids and having a molecular mass of ~ 16 kDa.
The Rat Leptin antagonist was mutated, resulting in L39A/D40A/F41A mutant.
The Rat Leptin antagonist is bound to 20 kDa mono-PEG at N-terminus, resulting in 35.6 kDa. The Rat Leptin triple anatagonist runs as a 48 kDa.
Leptin Antagonist Triple Mutant Rat Recombinant was purified by proprietary chromatographic techniques.

Product Specs

Description
Leptin Antagonist Triple Mutant Rat Recombinant is a single, non-glycosylated polypeptide chain containing 146 amino acids with an additional Alanine at the N-terminus. It has a molecular mass of approximately 16 kDa. This recombinant protein is a mutated form of Rat Leptin antagonist, featuring the mutations L39A/D40A/F41A. A 20 kDa mono-PEG molecule is attached to the N-terminus, resulting in a total molecular weight of 35.6 kDa. During SDS-PAGE analysis, the Rat Leptin triple antagonist appears as a 48 kDa band. The protein is purified using proprietary chromatographic techniques.
Physical Appearance
White, lyophilized powder.
Formulation
The Rat Leptin triple antagonist is lyophilized from a concentrated solution (0.65 mg/ml) containing 0.003 mM NaHCO3.
Solubility
To reconstitute the lyophilized Leptin Antagonist Triple Mutant Rat Recombinant, it is recommended to dissolve it in sterile water or sterile 0.4% NaHCO3 adjusted to a pH of 8-9. The final concentration should be at least 100 µg/ml. Following reconstitution, the solution can be further diluted with other aqueous solutions as needed.
Stability
Lyophilized Leptin Antagonist Triple Mutant Rat Recombinant remains stable at room temperature for several weeks. However, for long-term storage, it should be stored desiccated below -18°C. After reconstitution at concentrations greater than 0.1 mg/ml and in solutions containing up to 2 mM of the protein, and after filter sterilization, the solution can be stored at 4°C or room temperature for several weeks, making it suitable for long-term infusion studies using osmotic pumps. At lower concentrations, adding a carrier protein like 0.1% HSA or BSA is recommended. Avoid repeated freeze-thaw cycles.
Purity
The purity is determined to be greater than 99.0% using the following methods: (a) Gel filtration analysis, and (b) SDS-PAGE analysis.
Biological Activity
The Leptin Antagonist Triple Mutant Rat Recombinant exhibits a half-life of over 20 hours in circulation following subcutaneous injection. This antagonist effectively inhibits the proliferation of BAF/3 cells (stably transfected with the long form of the human leptin receptor) induced by leptin. While its in vitro activity is 5-6 fold lower compared to the non-pegylated antagonist, the pegylated form demonstrates a significant weight gain effect in vivo, primarily attributed to increased food intake.
Protein Content
Protein concentration is determined by measuring absorbance at 280 nm using a UV spectrophotometer. An extinction coefficient of 0.2 is employed for a 0.1% (1 mg/ml) solution at pH 8.0. This value is derived from the analysis of protein sequences using the PC GENE computer program (IntelliGenetics).
Source
Escherichia coli.

Product Science Overview

Leptin Antagonist Triple Mutant

The Leptin Antagonist Triple Mutant is a modified form of leptin designed to inhibit the action of natural leptin. This antagonist is particularly useful in research settings to study the effects of leptin inhibition. The triple mutant refers to specific mutations at three positions in the leptin protein: L39A, D40A, and F41A . These mutations significantly reduce the binding affinity of leptin to its receptor, thereby acting as an antagonist.

Pegylation

Pegylation is the process of attaching polyethylene glycol (PEG) chains to another molecule, in this case, the leptin antagonist. Pegylation is used to improve the pharmacokinetics of therapeutic proteins. It increases the stability and solubility of the protein, reduces immunogenicity, and prolongs the half-life in the bloodstream . For the Leptin Antagonist Triple Mutant, pegylation enhances its in vivo activity, making it more effective for research and potential therapeutic applications .

Rat Recombinant

The term rat recombinant indicates that the leptin antagonist is produced using recombinant DNA technology in a rat model. This involves inserting the gene encoding the leptin antagonist into a host organism, such as bacteria or yeast, to produce the protein in large quantities. The recombinant protein is then purified for use in research .

Applications and Significance

The Leptin Antagonist Triple Mutant Pegylated (Rat Recombinant) is primarily used in research to study obesity, metabolic disorders, and other conditions related to leptin signaling. By inhibiting leptin, researchers can better understand the hormone’s role in energy balance, appetite regulation, and body weight maintenance. This antagonist is also valuable in exploring potential therapeutic approaches for conditions like obesity and metabolic syndrome .

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