Recombinant Mouse Adiponectin

Recombinant mouse adiponectin is a valuable research tool with well-characterized biological activities and diverse applications across metabolic and tissue regeneration studies.

Metabolic and Glucose Homeostasis Research

Recombinant mouse adiponectin demonstrates robust effects on glucose metabolism and insulin sensitivity. The protein significantly reduces blood glucose levels in animal models and improves insulin signaling through activation of AMP-activated protein kinase (AMPK) and p38 mitogen-activated protein kinase (MAPK) pathways. In high-fat diet models, adiponectin gene therapy resulted in decreased weight gain, reduced fat accumulation, and improved glucose tolerance compared to control animals. The protein enhances insulin-stimulated phosphorylation of key signaling molecules including Akt and GSK-3β, making it particularly useful for studying hepatic insulin signaling and glucose homeostasis.

Lipid Metabolism and Energy Expenditure

Through AMPK and PPAR-alpha activation, recombinant adiponectin increases fatty acid transport, oxidation, and dissipation in skeletal muscle, leading to reduced intramyocellular lipid levels. Studies using this protein have demonstrated increased oxygen consumption and locomotor activity in treated animals, indicating enhanced whole-body energy expenditure. This makes it an excellent tool for investigating lipid metabolism and metabolic syndrome mechanisms.

Tissue Regeneration and Cell Biology

Recombinant adiponectin promotes proliferation and differentiation of tenocyte progenitor cells from diabetic patients when applied at concentrations of 10 μg/ml in vitro. The protein upregulates tenogenic gene expression including collagen I, collagen III, tenomodulin, and scleraxis, while maintaining selective differentiation pathways. This application is particularly valuable for studying diabetic tendinopathy and tissue repair mechanisms.

Experimental Versatility

The protein is available in multiple expression systems and formats suitable for various applications. Tested applications include cell culture and animal studies, Western blotting, ELISA, and bioassays. The availability of carrier-free formulations and high purity preparations (>95%) ensures minimal interference with downstream analyses.

You can use recombinant mouse adiponectin as a standard for quantification or calibration in your ELISA assays, provided that the recombinant protein is of high purity, its concentration is accurately known, and it is compatible with your assay’s antibodies and detection system.

Key considerations and supporting details:

• Recombinant Standards in Commercial Kits: Most commercial mouse adiponectin ELISA kits use recombinant mouse adiponectin as the standard for generating the calibration curve. These standards are typically highly purified and quantified, ensuring reliable calibration.

• Assay Compatibility: The recombinant standard must be recognized by the capture and detection antibodies used in your ELISA. Commercial kits validate their recombinant standards for this purpose, but if you are developing your own assay, confirm that your antibodies detect the recombinant form equivalently to the native protein.

• Standard Preparation: Prepare the standard curve by serially diluting the recombinant adiponectin in the same buffer or matrix as your samples to minimize matrix effects and ensure accurate quantification.

• Validation: If you are using a recombinant standard not supplied with a validated kit, it is best practice to validate its performance by assessing recovery, linearity, and parallelism with native samples. This ensures that the recombinant protein behaves similarly to endogenous adiponectin in your assay conditions.

• Documentation: Ensure the recombinant adiponectin’s source, expression system, and purity are well documented, as post-translational modifications or oligomerization state can affect antibody recognition and quantification accuracy.

Summary Table: Use of Recombinant Mouse Adiponectin as ELISA Standard

Conclusion:
Recombinant mouse adiponectin is widely accepted and routinely used as a standard for ELISA quantification, provided it meets the above criteria. Always validate its performance in your specific assay context for best results.

Recombinant Mouse Adiponectin has been validated for several key applications in published research, including cell culture, animal studies, Western blotting, ELISA, and bioassays.

Validated Applications:

• Cell Culture: Used to study adiponectin’s effects on cellular metabolism, proliferation, differentiation, and signaling pathways in various cell types.
• Animal Studies (In Vivo): Administered to mice to investigate its role in metabolic regulation, glucose homeostasis, insulin sensitivity, anti-inflammatory effects, cardiac protection, and tissue repair.
• Western Blotting: Utilized for detection and quantification of adiponectin protein expression in tissue or cell lysates.
• ELISA: Applied for quantitative measurement of adiponectin concentrations in biological samples such as serum, plasma, or cell culture supernatants.
• Bioassays: Used to assess the biological activity of adiponectin, including its effects on cell signaling, metabolic pathways, and functional outcomes in both cell-based and animal models.

Representative Research Applications:

• Metabolic Disease Models: Recombinant mouse adiponectin has been used to study its protective effects against obesity, diabetes, atherosclerosis, and metabolic syndrome in mouse models.
• Cardiovascular Research: Investigations include its role in preventing cardiac hypertrophy, improving cardiac function after myocardial infarction, and modulating inflammation.
• Tissue Repair and Regeneration: Shown to enhance proliferation and differentiation of progenitor cells, supporting tissue repair in conditions such as diabetic tendinopathy.
• Inflammation and Immune Modulation: Demonstrated anti-inflammatory effects in models of LPS-induced inflammation and kidney injury.
• Protein Detection and Quantification: Western blotting and ELISA protocols have been validated for adiponectin detection in various sample types.

Summary Table of Validated Applications

These applications are widely supported in the literature and are considered standard for recombinant mouse adiponectin research. If you require protocol details or specific experimental setups for any application, please specify the intended use.

To reconstitute and prepare Recombinant Mouse Adiponectin protein for cell culture experiments, dissolve the lyophilized protein in sterile, physiological buffer—commonly phosphate-buffered saline (PBS), pH 7.4, or sterile distilled water, depending on the manufacturer’s instructions. Use gentle mixing and avoid vigorous agitation or foaming to preserve protein integrity.

Step-by-step protocol:

• Equilibrate the vial to room temperature before opening.
• Add sterile PBS or distilled water to the vial. The reconstitution volume depends on the desired final concentration and the amount of protein in the vial. For example, add 1 mL for a typical standard preparation, but consult the product datasheet for exact volume.
• Mix gently by inverting or slow pipetting. Do not vortex or shake vigorously, as this can denature the protein and cause foaming.
• Allow the solution to sit at room temperature for 10–30 minutes with gentle agitation to ensure complete dissolution.
• Aliquot the reconstituted protein to avoid repeated freeze-thaw cycles, which can degrade the protein.
• Store aliquots at −20°C or −80°C for long-term storage. For short-term use, keep at 4°C and use within a few days.
• Prepare working dilutions in cell culture medium or PBS containing at least 0.1% BSA to minimize adsorption and loss of activity. Filter-sterilize if necessary.

Additional notes for cell culture:

• Always use sterile technique to prevent contamination.
• If the protein contains stabilizers (e.g., trehalose, BSA), verify compatibility with your cell culture system.
• For functional assays, optimize the concentration for your specific cell type and experimental design. Typical working concentrations range from 10 ng/mL to 1 μg/mL, but titration is recommended.
• If using in serum-free media, supplement with carrier protein (e.g., BSA) to stabilize adiponectin.

Summary Table:

Always consult the specific product datasheet for any unique requirements or recommended buffers for your recombinant mouse adiponectin preparation.