Recombinant Mouse Resistin

Recombinant Mouse Resistin is widely used in research applications to investigate its roles in metabolism, inflammation, and host defense, particularly as a model for studying insulin resistance, adipogenesis, and immune modulation in murine systems.

Key scientific reasons to use recombinant mouse resistin include:

• Modeling Insulin Resistance and Metabolic Disease: Mouse resistin is a critical factor in the development of insulin resistance and impaired glucose metabolism. Administration of recombinant mouse resistin to rodents acutely impairs hepatic insulin sensitivity and increases glucose production, making it a valuable tool for studying the mechanisms linking obesity to diabetes and metabolic syndrome. It is also used to test interventions that may reverse or exacerbate these effects.

• Adipogenesis and Glucose Uptake Studies: Recombinant mouse resistin modulates adipocyte differentiation and glucose uptake, acting as an inhibitory ligand for receptors such as ROR1. It is used to dissect the molecular pathways involved in adipogenesis and the development of insulin resistance in cell culture models like 3T3-L1 preadipocytes.

• Immunomodulatory and Proinflammatory Effects: Mouse resistin induces the expression of proinflammatory cytokines (e.g., IL-6) in adipocytes and immune cells, and promotes the secretion of other inflammatory mediators. This makes it useful for studying the links between metabolic dysfunction and inflammation, as well as the cellular signaling pathways involved.

• Innate Immunity and Antimicrobial Activity: Recent research highlights resistin’s role as a host defense peptide, capable of directly killing bacteria and modulating immune responses. Recombinant mouse resistin can be used to study its antimicrobial properties and its effects on immune cell recruitment, cytokine release, and formation of neutrophil extracellular traps (NETs).

• Bioassays and ELISA Standards: Recombinant mouse resistin is used as a standard in ELISA assays for quantifying resistin levels in biological samples, and in bioassays to assess its biological activity in vitro.

• Species-Specific Mechanistic Studies: Mouse resistin shares partial sequence identity with human and rat resistin, but exhibits distinct biological activities, such as not promoting lipolysis in adipocytes. Using recombinant mouse resistin allows for species-specific mechanistic studies relevant to murine models.

Typical applications include:

• In vivo administration to rodents for metabolic and immunological studies.
• In vitro cell culture experiments to assess effects on adipogenesis, cytokine production, and immune cell activation.
• Use as a standard in immunoassays.

In summary, recombinant mouse resistin is essential for dissecting the molecular and physiological roles of resistin in murine models of metabolic disease, inflammation, and innate immunity, providing a controlled and reproducible reagent for mechanistic and translational research.

Yes, recombinant mouse Resistin can be used as a standard for quantification or calibration in ELISA assays, provided it is of high purity, correctly quantified, and compatible with your assay system. This is a common practice in ELISA development and commercial kits, where recombinant proteins are routinely used to generate standard curves for quantification of target analytes.

Key considerations and supporting details:

• Recombinant mouse Resistin is widely used as an ELISA standard: Commercial ELISA kits for mouse Resistin typically use recombinant mouse Resistin as the calibrator to generate standard curves for quantification in serum, plasma, and cell culture samples.
• Purity and quantification: The recombinant protein should be of high purity (typically >80–95%) and accurately quantified, as impurities or inaccurate concentration will affect the reliability of your standard curve.
• Formulation: Some recombinant proteins are supplied with carrier proteins (e.g., BSA) to enhance stability, while others are carrier-free. Choose the formulation that best matches your assay requirements and does not interfere with detection.
• Validation: Ensure that the recombinant Resistin standard is recognized by the capture and detection antibodies in your ELISA. Most commercial kits validate their standards for this purpose, but if you are developing your own assay, you should confirm cross-reactivity and linearity.
• Standard curve preparation: Prepare a serial dilution of the recombinant Resistin in the same buffer or matrix as your samples to minimize matrix effects and ensure accurate quantification.
• Documentation: Refer to the technical datasheet or manual for your specific recombinant protein or ELISA kit for recommended concentration ranges and reconstitution protocols.

Limitations and best practices:

• If your ELISA is designed for native Resistin from biological samples, confirm that the recombinant standard mimics the native protein’s conformation and epitopes, as post-translational modifications or folding differences can affect antibody recognition.
• Always validate the standard curve in your specific assay context, especially if using a recombinant standard from a different source or expression system than the kit manufacturer recommends.

Summary:
Recombinant mouse Resistin is suitable and commonly used as a standard for ELISA quantification, provided it is validated for your assay and prepared according to best practices for standard curve generation.

Recombinant Mouse Resistin has been validated for several applications in published research, primarily in the study of metabolic regulation, inflammation, and innate immunity.

Key validated applications include:

• Bioassays: Used to assess biological activity, such as induction of proinflammatory molecules in adipocytes, promotion of hepatic gluconeogenesis, and insulin resistance in cell-based systems. For example, recombinant mouse resistin has been shown to induce IL-6 expression in 3T3-L1 cells in a dose-dependent manner.

• ELISA Standard: Employed as a standard in enzyme-linked immunosorbent assays (ELISA) for quantifying resistin levels in biological samples.

• Cellular and Molecular Mechanism Studies: Utilized in mechanistic studies to investigate its interaction with receptors (e.g., ROR1), modulation of signaling pathways (such as ERK1/2 and SOCS-3), and effects on glucose uptake and adipogenesis in 3T3-L1 preadipocytes.

• In Vivo Functional Studies: Used in animal models to restore circulating resistin levels and study its effects on insulin resistance, glucose metabolism, and inflammatory responses.

• Antimicrobial and Immunomodulatory Assays: Applied in studies exploring its role as a host defense peptide, including direct antimicrobial activity and modulation of immune cell recruitment, cytokine release, and neutrophil extracellular trap (NET) formation.

Supporting details and representative published uses:

• Bioassay: Recombinant mouse resistin has been used to induce proinflammatory molecule production in adipocytes and to study its effects on hepatic gluconeogenesis and insulin resistance.
• ELISA Standard: Validated as a standard for quantifying resistin in ELISA-based assays.
• Adipogenesis and Glucose Uptake: Used to modulate adipogenesis and glucose uptake in 3T3-L1 cells, and to study receptor interactions (e.g., ROR1).
• In Vivo Infusion: Infused into mice to study acute effects on circulating resistin levels and metabolic phenotypes.
• Immunological Studies: Used to investigate its antimicrobial properties and immunomodulatory functions, including effects on cytokine production and immune cell activation.

These applications are supported by both primary research articles and product validation data from multiple sources.

Reconstitution Protocol

Initial Preparation

Begin by centrifuging the lyophilized vial at low speed (3000-3500 rpm) for 5 minutes before opening. This concentrates any protein that may have adhered to the cap or tube walls during shipping. The recommended reconstitution concentration is 250 μg/mL in sterile PBS, though alternative concentrations of 0.1-1.0 mg/mL are also acceptable depending on your experimental requirements.

Dissolution Steps

When reconstituting, add the appropriate volume of sterile PBS slowly to the lyophilized powder. Do not vortex the solution, as vigorous mixing can break chemical bonds and inactivate the protein. Instead, gently shake the vial or use a pipette to blow gently several times. Allow approximately 20 minutes at room temperature for complete dissolution.

Formulation Considerations

Carrier Protein Selection

For cell and tissue culture applications, use the formulation containing bovine serum albumin (BSA) as a carrier protein. Reconstitute this version at 250 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin. The carrier protein serves multiple critical functions: it enhances protein stability, increases shelf-life, prevents protein adsorption to plastic tube walls, and allows storage at more dilute concentrations.

If your experimental design requires the absence of BSA—for instance, in certain immunoassays or binding studies where BSA could interfere—use the carrier-free formulation reconstituted in sterile PBS alone.

Storage and Stability

Short-term Storage

After initial reconstitution, the protein can be stored at 4°C for up to 1 week. For working aliquots used in cell culture, maintain a minimum volume of at least 20 μL to minimize protein loss from adsorption to container walls.

Long-term Storage

For extended storage beyond one week, further dilute the reconstituted protein in a buffer containing carrier proteins (0.1% BSA, 10% fetal bovine serum, 5% human serum albumin, or 5% trehalose) and divide into separate aliquots. Store these working aliquots at -20°C to -80°C for 3-6 months. Use a manual defrost freezer and avoid repeated freeze-thaw cycles, as each cycle decreases protein activity.

The lyophilized powder itself remains stable at -20°C for 12 months from the date of receipt.

Key Practical Considerations

Ensure all reconstitution buffers and equipment are sterile for cell culture applications. When preparing dilutions for experiments, use appropriate dilution buffers (such as culture medium or PBS) that also contain carrier proteins or stabilizers to maintain protein integrity. Always refer to the product Certificate of Analysis for any specific recommendations tailored to your particular lot, as formulation details may vary slightly between preparations.