Recombinant Mouse gAcrp30 (Adipocyte Complement-Related Protein)

Recombinant Mouse gAcrp30 (Adipocyte Complement-Related Protein, globular adiponectin) is used in research to study metabolic regulation, insulin sensitivity, lipid metabolism, and energy homeostasis, as it is a biologically active fragment of adiponectin with distinct and potent physiological effects.

Key reasons to use recombinant mouse gAcrp30 in research applications:

• Metabolic Regulation: gAcrp30 is a proteolytic cleavage product of full-length adiponectin and retains the biologically active globular domain, which is particularly effective in stimulating fatty acid oxidation in muscle and reducing plasma free fatty acids. This makes it a valuable tool for studying mechanisms of lipid metabolism and energy expenditure.

• Insulin Sensitization: gAcrp30 has been shown to improve insulin sensitivity and decrease hyperglycemia, making it relevant for research into diabetes, obesity, and metabolic syndrome. Its expression is reduced in insulin-resistant and obese states, and increasing its levels can enhance whole-body insulin sensitivity.

• Weight Regulation: In vivo studies demonstrate that administration of gAcrp30 leads to significant and sustained weight loss in mice on high-fat diets, without affecting food intake, by increasing peripheral fatty acid oxidation rather than altering appetite.

• Distinct Receptor Interactions: The globular domain (gAcrp30) interacts with adiponectin receptors (AdipoR1 and AdipoR2) with different affinities compared to full-length adiponectin, allowing researchers to dissect receptor-specific signaling pathways in muscle and liver tissues.

• Anti-inflammatory and Anti-atherogenic Effects: Like full-length adiponectin, gAcrp30 exhibits anti-inflammatory and anti-atherogenic properties, relevant for cardiovascular and metabolic disease models.

• Experimental Versatility: Recombinant gAcrp30 is available in carrier-free and highly purified forms, suitable for cell culture, in vivo administration, and biochemical assays, enabling controlled mechanistic studies without confounding factors such as carrier proteins or endotoxins.

• Modeling Human Disease: Mouse gAcrp30 shares high sequence identity with the human protein, making it a suitable model for translational research into human metabolic disorders.

Summary of applications:

• Investigating mechanisms of insulin resistance and diabetes
• Studying lipid metabolism and energy homeostasis
• Exploring anti-inflammatory and cardiovascular protective pathways
• Testing pharmacological interventions for obesity and metabolic syndrome

Best practices: Use carrier-free preparations for receptor binding or signaling studies where BSA or other carriers may interfere, and ensure low endotoxin levels for in vivo or cell-based assays.

In summary, recombinant mouse gAcrp30 is a powerful tool for dissecting the molecular and physiological roles of adiponectin in metabolism, obesity, diabetes, and related fields.

You can use recombinant Mouse gAcrp30 (globular Adipocyte Complement-Related Protein) as a standard for quantification or calibration in ELISA assays, provided the assay is designed to detect the globular domain and the recombinant protein is well-characterized and pure. However, there are important technical considerations:

• Assay Compatibility: ELISA kits for mouse Adiponectin/Acrp30 typically use standards based on the full-length protein, not just the globular domain (gAcrp30). If your ELISA is specific for the globular domain, recombinant gAcrp30 is appropriate. If your assay detects full-length Adiponectin, using gAcrp30 may result in inaccurate quantification due to differences in epitope recognition and molecular weight.

• Standard Curve Preparation: For quantitative ELISA, a standard curve must be prepared using a purified protein standard that matches the analyte detected by the assay. Recombinant gAcrp30 can be used if it is pure, its concentration is accurately determined, and it is reconstituted and handled according to best practices for ELISA standards.

• Protein Form and Purity: Ensure the recombinant gAcrp30 is of high purity (typically >95%), with minimal endotoxin and verified identity (e.g., by SDS-PAGE and mass spectrometry). Carrier-free preparations are preferred for ELISA standards to avoid interference from stabilizers like BSA.

• Calibration and Validation: If switching from a full-length Adiponectin standard to gAcrp30, you must validate the assay for linearity, recovery, and sensitivity using the recombinant protein to ensure accurate quantification. The standard curve should cover the expected concentration range in your samples.

Summary of best practices:

• Use recombinant gAcrp30 only if your ELISA detects the globular domain.
• Confirm the purity and concentration of the recombinant protein.
• Prepare and validate the standard curve according to ELISA guidelines.
• If your assay is designed for full-length Adiponectin, use a matching standard for accurate results.

If you provide more details about your ELISA format (capture/detection antibodies, target domain), more specific recommendations can be given.

Recombinant Mouse gAcrp30 (Adipocyte Complement-Related Protein, globular adiponectin) has been validated for several key applications in published research, primarily in studies of metabolic regulation, cell signaling, and disease models.

Validated Applications:

• In Vivo Studies:
  Used to investigate metabolic effects, such as insulin sensitization, anti-diabetic activity, and anti-atherogenic properties in mouse models. For example, recombinant gAcrp30 has been administered to mice to study its impact on insulin sensitivity, glucose metabolism, and weight loss.

• Cell-Based Bioassays:
  Applied to cultured cells to assess biological activity, including inhibition of cell proliferation (e.g., mouse M1 myeloid cells), modulation of signaling pathways, and effects on hormone secretion (e.g., inhibition of GnRH secretion from hypothalamic neurons).

• ELISA Standard:
  Utilized as a standard protein in ELISA assays for quantification of adiponectin/gAcrp30 levels in biological samples.

Representative Published Research:

• Metabolic Regulation:
  Recombinant gAcrp30 has been used to demonstrate enhancement of hepatic insulin action and whole-body glucose metabolism, supporting its role as a potent insulin sensitizer.

• Obesity and Diabetes Models:
  Studies have validated its use in obese and diabetic mouse models to show increased plasma gAcrp30 levels following treatment with PPARγ agonists, linking it to improved insulin sensitivity and suggesting its utility as a biomarker for PPARγ activation.

• Cell Signaling and Proliferation:
  Research has shown that recombinant gAcrp30 can block leptin-induced cell proliferation and modulate multiple signaling cascades in colon epithelial cells.

• Neuroendocrine Regulation:
  It has been used to inhibit GnRH secretion in hypothalamic neurons, indicating a role in neuroendocrine signaling.

Summary Table of Validated Applications

Additional Notes:

• The recombinant globular domain (gAcrp30) is often preferred for studies targeting AdipoR1 receptor signaling, as it has higher affinity for this receptor compared to full-length adiponectin.
• Published research supports its use in both mouse and human systems for mechanistic studies of metabolic disease, obesity, and insulin resistance.

If you require specific protocols or further details on experimental design, please specify the application area.

To reconstitute and prepare Recombinant Mouse gAcrp30 (Adipocyte Complement-Related Protein) for cell culture experiments, dissolve the lyophilized protein in sterile distilled water or buffer to a concentration between 0.1–1.0 mg/mL, then aliquot and store at −20 °C with a carrier protein (such as BSA or HSA) to maintain stability and avoid repeated freeze-thaw cycles.

Detailed protocol and best practices:

• Reconstitution:

  • Centrifuge the vial briefly to collect the powder at the bottom.
  • Add sterile distilled water or buffer (e.g., PBS) to achieve the desired concentration, typically 0.1–1.0 mg/mL.
  • Gently pipette up and down or vortex briefly to fully dissolve the protein. Avoid vigorous agitation to prevent denaturation.
  • If the datasheet specifies a particular buffer or pH, follow those instructions. Some protocols recommend adding 0.1% BSA or HSA as a carrier protein to enhance stability, especially for cell culture applications.
  • Allow the solution to sit at room temperature for a few minutes to ensure complete dissolution.

• Aliquoting and Storage:

  • Aliquot the reconstituted protein into working volumes (e.g., 10–100 µL) to minimize freeze-thaw cycles.
  • Store aliquots at −20 °C for up to 3 months; for short-term use, storage at 4 °C is acceptable for a few days.
  • If using a carrier protein, stability is improved and aggregation is minimized.
  • Avoid repeated freeze-thaw cycles, as this can lead to protein degradation and loss of activity.

• Preparation for Cell Culture:

  • Thaw aliquots on ice and dilute to the desired working concentration in cell culture medium immediately before use.
  • Filter-sterilize the final working solution if sterility is required and the protein is not already supplied sterile.
  • Typical working concentrations for cell culture range from 5–20 µg/mL, but optimize based on your experimental design and cell type.

Additional notes:

• If the recombinant protein is supplied as a solution (not lyophilized), check the datasheet for recommended storage and handling. Carrier-free formulations may require special attention to prevent adsorption to plasticware.
• Always consult the specific product datasheet for any unique instructions regarding buffer composition, pH, or additives.

Summary Table:

Following these guidelines will help ensure the stability and biological activity of recombinant mouse gAcrp30 for cell culture experiments.