Recombinant Mouse Leptin R

Using Recombinant Mouse Leptin R (Leptin Receptor, LepR) in research applications is essential for studying the molecular mechanisms of leptin signaling, energy homeostasis, metabolic regulation, and related disease models in mice. This recombinant protein enables precise in vitro and in vivo investigations of leptin–receptor interactions, downstream signaling pathways, and the physiological or pathological roles of leptin in metabolism, immunity, and neuroendocrine function.

Key scientific applications and rationale include:

• Mechanistic Studies of Leptin Signaling: Recombinant Mouse Leptin R allows researchers to dissect how leptin binds and activates its receptor, facilitating studies of receptor structure, ligand specificity, and downstream signaling cascades such as JAK/STAT, PI3K, and MAPK pathways.
• Modeling Metabolic and Endocrine Disorders: Since mutations or dysregulation of LepR cause severe obesity and metabolic dysfunction in mice, using recombinant LepR helps model and understand diseases like obesity, diabetes, and lipodystrophy, and test therapeutic interventions.
• Receptor–Ligand Binding Assays: Recombinant LepR is critical for in vitro binding assays, screening for leptin analogs, antagonists, or agonists, and characterizing the affinity and kinetics of leptin–receptor interactions.
• Immunological and Inflammatory Research: LepR is involved in immune modulation and inflammatory responses. Recombinant LepR can be used to study leptin’s effects on immune cells, cytokine production, and inflammation in mouse models.
• Development of Diagnostic and Therapeutic Tools: Soluble forms of recombinant LepR can serve as decoys to neutralize leptin activity, or as standards in ELISA and other immunoassays for quantifying leptin or its receptor in biological samples.
• Cancer and Cardiovascular Research: LepR expression is altered in various diseases, including cancer and heart failure. Recombinant LepR enables investigation of leptin’s role in tumor biology, angiogenesis, and cardiovascular pathology.

Best practices for using recombinant proteins include validating activity, using appropriate controls, and matching the recombinant receptor’s isoform and post-translational modifications to the biological context of your study.

In summary, Recombinant Mouse Leptin R is a versatile tool for elucidating leptin biology, modeling disease, and developing new diagnostic and therapeutic strategies in mouse-based research.

Recombinant Mouse Leptin R (Leptin Receptor) can be used as a standard for quantification or calibration in ELISA assays, provided the assay is validated to recognize both natural and recombinant forms.

Most commercial Mouse Leptin R ELISA kits are specifically designed to quantify both natural and recombinant Mouse Leptin R in biological samples such as serum, plasma, and cell culture supernatants. The standard curve in these kits is typically generated using recombinant Mouse Leptin R, and the assay antibodies are validated for specificity to both forms. This ensures that the quantification of endogenous (natural) Leptin R in your samples is accurate when calibrated against the recombinant standard.

Key considerations for using recombinant Mouse Leptin R as a standard:

• Assay validation: Confirm that your ELISA kit or custom assay recognizes both recombinant and natural Mouse Leptin R with parallel dose-response curves. Most reputable kits provide this validation in their documentation.
• Standard curve preparation: Prepare a dilution series of recombinant Mouse Leptin R in the same matrix as your samples (e.g., assay buffer or calibrator diluent) to generate the standard curve.
• Quantification: Sample concentrations are determined by interpolation from the standard curve generated with recombinant Mouse Leptin R.
• Sensitivity and range: Ensure the concentration range of your recombinant standard matches the expected levels in your samples. For example, typical standard curve ranges are 3.13–200 ng/mL or similar, with sensitivities down to 0.039 ng/mL depending on the kit.

Limitations:

• The recombinant standard should be of high purity and properly quantified.
• The assay should be for research use only, not for diagnostic procedures.

Summary Table: Use of Recombinant Mouse Leptin R in ELISA Calibration

If you are developing a custom ELISA, ensure your capture and detection antibodies are validated for both recombinant and natural Mouse Leptin R, and always run parallelism tests to confirm equivalence.

Recombinant Mouse Leptin Receptor (Leptin R) has been validated for several key applications in published research, primarily in bioassays, inhibition studies, and as a tool for investigating leptin signaling in vitro and in vivo.

Validated Applications

• Inhibition Bioassays:
  The recombinant mouse Leptin R Fc chimera protein has been validated for use in bioassays measuring its ability to inhibit leptin-dependent proliferation of BaF3 mouse pro-B cells transfected with human Leptin R. This is a standard functional assay to confirm receptor activity and specificity.

• Detection and Quantification:
  Recombinant Leptin R is used as a standard or capture reagent in ELISA and other immunoassays to detect leptin or leptin receptor interactions.

• Cell Culture Studies:
  It is applied in cell-based assays to study leptin signaling, receptor binding, and downstream effects in various cell types, including hematopoietic and immune cells.

• In Vivo Studies:
  Recombinant mouse leptin and its receptor have been used in animal models to investigate physiological roles such as body weight regulation, hematopoiesis, reproduction, and metabolic disease mechanisms. These studies often involve administration of recombinant proteins to mice and assessment of phenotypic or molecular outcomes.

• Therapeutic and Mechanistic Research:
  The receptor is utilized in research exploring therapeutic strategies for obesity, metabolic syndrome, neurodegenerative diseases (e.g., ALS), and immune modulation.

Supporting Details

• Functional Validation:
  The ability of recombinant Leptin R to bind leptin and inhibit its biological activity is a critical validation step, often performed using proliferation assays in engineered cell lines.

• Immunological and Metabolic Studies:
  Recombinant Leptin R is used to dissect leptin’s role in immune cell metabolism, inflammation, and autoimmunity, as well as its effects on glucose homeostasis and neuroprotection in mouse models.

• Assay Development:
  The protein serves as a reagent for developing and validating immunoassays (e.g., ELISA, mass spectrometry) for leptin and its receptor, enabling quantification in biological samples.

Summary Table

In summary, recombinant mouse Leptin R is a validated tool for bioassays, immunoassays, cell culture, and in vivo studies, supporting research into leptin biology, metabolic regulation, and therapeutic development.

Reconstitution Protocol

Recombinant Mouse Leptin R protein should be reconstituted at a concentration of 100 μg/mL in phosphate-buffered saline (PBS). Before opening the vial, briefly centrifuge it to bring the contents to the bottom, which helps ensure complete recovery of the lyophilized protein.

When reconstituting, gently pipette the PBS solution down the sides of the vial rather than directly onto the protein cake. This gentle approach minimizes foaming and protein denaturation. Allow the protein to dissolve gradually by gentle mixing rather than vigorous agitation.

Storage and Stability

After aseptic reconstitution, store the protein at 2–8°C for up to one month, or at −20°C to −70°C for longer-term storage in a manual defrost freezer. Avoid repeated freeze-thaw cycles, as these can compromise protein integrity and biological activity. If you anticipate multiple uses, consider preparing aliquots to minimize freeze-thaw exposure.

Cell Culture Preparation

For cell culture experiments, the reconstituted stock solution can be further diluted to your desired working concentration using appropriate culture media or buffers. The specific dilution will depend on your experimental design and the biological responses you are measuring. Ensure all solutions used for dilution are sterile and appropriate for your cell culture system.

Key Considerations

• Use sterile technique throughout the reconstitution and preparation process
• Allow adequate time for complete dissolution before use
• Document the reconstitution date and store information for traceability
• Verify protein concentration if quantitative analysis is required for your experiments