Leptin Mouse, PEG

Pegylated Mouse Leptin Recombinant
Cat. No.
BT20961
Source
Escherichia Coli.
Synonyms
OB Protein, Obesity Protein, OBS, Obesity factor.
Appearance
Sterile Filtered White lyophilized (freeze-dried) powder.
Purity
Greater than 99.0% as determined by:
(a) Analysis by Gel-Filtration.
(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 Mono-Pegylated Mouse Recombinant produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 146 amino acids and an additional Ala at N-terminus. Pegylated Mouse Leptin contains PEG 20 kDa at its N-terminus and having a molecular mass of 35.6 kDa as determined by mass spectrometry.
Since its enlarged hydrodymanic volume Pegylated Leptin runs on SDS-PAGE as A 48 kDa protein and in gel-filtration on Superdex 200 as over 100 kDa protein.
Pegylated Mouse Leptin half-life in circulation after SC injection was over 20 hours. Mouse Leptin was purified by proprietary chromatographic techniques according to Salomon et al (2006) Protein Expression and Purification 47, 128–136 and then pegylated.

Product Specs

Introduction
Leptin, a 16-kDa hormone produced by adipocytes, plays a crucial role in regulating energy balance and appetite. It acts as a signal from fat cells, informing the brain about the body's energy stores.
Description
This product consists of mono-pegylated recombinant mouse leptin. Produced in E. coli, it is a single, non-glycosylated polypeptide chain of 146 amino acids with an additional alanine at the N-terminus. The PEGylation, achieved with a 20 kDa PEG molecule at the N-terminus, results in a 35.6 kDa protein as determined by mass spectrometry. Notably, the pegylation affects its apparent size in SDS-PAGE (48 kDa) and gel filtration (over 100 kDa) due to its increased hydrodynamic volume. This modification extends the protein's half-life in circulation to over 20 hours following subcutaneous injection. The purification process involves proprietary chromatographic methods as outlined in Salomon et al. (2006) Protein Expression and Purification 47, 128–136, followed by the pegylation step.
Physical Appearance
The product appears as a white, lyophilized powder that has undergone sterile filtration.
Formulation
The lyophilized mouse leptin is prepared from a 0.65 mg/ml solution containing 0.003 mM sodium bicarbonate.
Solubility
For reconstitution, dissolve the lyophilized leptin in sterile 0.4% sodium bicarbonate solution adjusted to pH 8.5. Aim for a minimum concentration of 100 µg/ml. This solution can be further diluted in other aqueous solutions as needed.
Stability
While the lyophilized leptin remains stable at room temperature for up to three weeks, it is recommended to store it desiccated at -18°C or below. After reconstitution, store at 4°C for 2-7 days. For long-term storage, freeze at -18°C or below after adding a carrier protein like HSA or BSA (0.1%). Avoid repeated freeze-thaw cycles.
Purity
The purity is greater than 99.0% as confirmed by gel filtration and SDS-PAGE analyses.
Biological Activity
This pegylated mouse leptin effectively stimulates the proliferation of BAF/3 cells expressing the long form of the human leptin receptor. Although its in vitro activity is slightly lower compared to the non-pegylated form, pegylated mouse leptin demonstrates a significantly stronger weight-reducing effect in vivo, primarily due to its appetite-suppressing properties.
Synonyms
OB Protein, Obesity Protein, OBS, Obesity factor.
Source
Escherichia Coli.

Product Science Overview

Definition and Classification

Pegylated Mouse Leptin Recombinant is a modified form of leptin, a 16-kDa peptide hormone secreted by adipose tissue. Leptin plays a crucial role in regulating food intake and energy balance. The pegylation process involves attaching polyethylene glycol (PEG) to the leptin molecule, enhancing its stability and half-life in circulation .

Biological Properties

Pegylated Mouse Leptin Recombinant is a single, non-glycosylated polypeptide chain containing 146 amino acids, with an additional alanine at the N-terminus. The PEG moiety, typically 20 kDa, is attached to the N-terminus, resulting in a molecular mass of 35.6 kDa as determined by mass spectrometry . This modification increases the hydrodynamic volume, causing the protein to run as a 48 kDa protein on SDS-PAGE and over 100 kDa in gel-filtration on Superdex 200 .

Expression Patterns and Tissue Distribution

Leptin is primarily expressed in white adipose tissue, but it can also be found in other tissues such as the stomach, placenta, and skeletal muscle. The recombinant form is produced in Escherichia coli using proprietary chromatographic techniques .

Biological Functions

Leptin is a key regulator of energy homeostasis, providing feedback signals from fat cells to the brain to control body fat stores. It influences various physiological processes, including appetite regulation, energy expenditure, and glucose metabolism .

Modes of Action

Leptin exerts its effects by binding to the leptin receptor (LEPR), a class I cytokine receptor. This interaction activates the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway, leading to the regulation of gene expression involved in energy balance .

Regulatory Mechanisms

The pegylation of leptin significantly enhances its half-life in circulation, making it more effective in reducing food intake and body weight in vivo compared to non-pegylated leptin . This modification also improves the bioavailability and stability of the protein, allowing for more sustained biological activity .

Applications and Research

Pegylated Mouse Leptin Recombinant is widely used in laboratory research to study obesity, diabetes, and other metabolic disorders. Its ability to stimulate the proliferation of BAF/3 cells stably transfected with the long form of the human leptin receptor makes it a valuable tool for investigating leptin signaling pathways .

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