Recombinant Mouse IGF-II

Recombinant Mouse IGF-II is used in research applications to study its roles in cell growth, differentiation, survival, and neuroprotection, particularly in mouse models relevant to developmental biology, neuroscience, and disease modeling.

Key scientific reasons to use recombinant Mouse IGF-II include:

• Modeling Mouse Physiology: Mouse IGF-II is species-specific and closely mimics endogenous mouse protein, making it ideal for in vitro and in vivo studies in mouse systems, ensuring physiological relevance for developmental, metabolic, or disease research.
• Neural Development and Neuroprotection: IGF-II is critical for nervous system proliferation, differentiation, myelination, and neuroprotection. It has been shown to promote neurogenesis, synaptogenesis, and protect against neurodegenerative insults in mouse models, including Alzheimer’s and Parkinson’s disease.
• Cell Proliferation and Differentiation: IGF-II stimulates proliferation and migration of various cell types, supporting studies in embryonic development, tissue regeneration, and stem cell biology.
• Disease Modeling: Recombinant Mouse IGF-II is used to investigate mechanisms underlying diseases such as neurodegeneration, cancer, and metabolic disorders, and to test therapeutic strategies targeting IGF-II pathways.
• Receptor-Specific Studies: Mouse IGF-II interacts with IGF1R and IGF2R, allowing detailed analysis of receptor-mediated signaling pathways in mouse cells and tissues.

In summary, recombinant Mouse IGF-II is a versatile tool for dissecting IGF-II’s biological functions in mouse models, enabling translational research in development, neurobiology, and disease.

Yes, recombinant Mouse IGF-II can be used as a standard for quantification or calibration in ELISA assays, provided it is validated for this purpose and matches the assay’s requirements. Recombinant proteins are commonly used as standards in ELISA kits to generate calibration curves for quantifying target analytes.

Key considerations for use:

• Validation: The recombinant Mouse IGF-II must be validated for use as a standard in your specific ELISA format. Many commercial ELISA kits for mouse IGF-II use recombinant IGF-II as their standard, demonstrating its suitability for calibration.
• Parallelism: The standard curve generated with recombinant IGF-II should be parallel to curves obtained with natural samples, ensuring accurate quantification.
• Purity and Formulation: Use carrier-free or BSA-formulated recombinant IGF-II as appropriate for your assay type. Carrier-free forms are preferred for ELISA standards to avoid interference.
• Concentration Range: Prepare the standard curve within the assay’s validated range (e.g., 2.3–150 ng/mL for some kits).
• Matrix Effects: If your samples are in complex matrices (e.g., serum, plasma), confirm that the recombinant standard behaves similarly to endogenous IGF-II in those matrices.

Protocol best practices:

• Reconstitute and dilute the recombinant IGF-II according to the ELISA kit instructions.
• Run standards in duplicate or triplicate for accuracy.
• Include both low and high controls in each assay run.
• Confirm that the recombinant standard is recognized by the antibodies used in your ELISA (most commercial kits specify compatibility with both natural and recombinant IGF-II).

Limitations:

• Not all recombinant proteins are suitable for all ELISA formats; always check compatibility with your specific kit and antibody pair.
• If using a custom or in-house ELISA, additional validation may be required to ensure the recombinant standard’s equivalence to native IGF-II.

Summary Table: Recombinant Mouse IGF-II as ELISA Standard

*Validation required for custom assays.

In conclusion, recombinant Mouse IGF-II is widely accepted as a standard for ELISA quantification, provided it is validated for your assay system and matches the kit’s specifications.

Recombinant Mouse IGF-II has been validated in published research for several key applications, primarily in studies of bioactivity, neuroprotection, neurodevelopment, metabolic regulation, and cell signaling.

Validated Applications in Published Research:

• Bioactivity Assays: Recombinant Mouse IGF-II is routinely used to assess its biological activity in vitro, including cell proliferation and differentiation assays.
• Neuroprotection and Neurodegeneration Models: IGF-II has been applied in rodent models of Alzheimer's disease (AD), Huntington's disease (HD), and Parkinson's disease (PD) to evaluate its neuroprotective effects, such as reducing amyloid β plaques, protecting neurons from neurotoxicity, and improving cognitive function.
• Memory and Cognitive Function Studies: Direct injection or overexpression of IGF-II in the hippocampus or CNS has been shown to enhance memory, reverse cognitive deficits, and promote synaptic plasticity in animal models.
• Cell Signaling and Differentiation: IGF-II is used to study signaling pathways involved in tissue differentiation, muscle development, and glucose metabolism in adipose tissue, skeletal muscle, and liver.
• Endothelial Cell Migration and Angiogenesis: IGF-II promotes endothelial cell migration and tube formation, supporting its use in angiogenesis and vascular biology research.
• Functional Assays: It has been used in functional assays to assess its role in myogenic differentiation, regulation of transcription factors, and metabolic processes.
• ELISA Standard and Western Blot: Recombinant Mouse IGF-II serves as a standard in ELISA development and is detected in Western blot assays for protein quantification and characterization.
• Cell Culture Supplementation: It is used as a supplement in cell culture to support growth, differentiation, and survival of various cell types.

Representative Published Research Applications:

• Alzheimer’s Disease: IGF-II administration improved cognitive function, reduced amyloid pathology, and stimulated neurogenesis in rodent AD models.
• Huntington’s Disease: IGF-II gene therapy decreased mutant huntingtin aggregation and protected neurons in HD models.
• Autism Spectrum Disorders: IGF-II reversed social interaction deficits and improved social memory in mouse models.
• Angiogenesis: IGF-II and its variants enhanced endothelial migration and tube formation in vitro.
• Muscle Differentiation: IGF-II positively regulated myogenic transcription factors and muscle terminal differentiation.

Summary Table:

These applications are supported by multiple peer-reviewed studies and technical validations, demonstrating the versatility of recombinant Mouse IGF-II in both basic and translational research contexts.

To reconstitute and prepare Recombinant Mouse IGF-II protein for cell culture experiments, dissolve the lyophilized protein in sterile water or phosphate-buffered saline (PBS), typically to a concentration of at least 0.1–0.2 mg/mL, and incubate at room temperature for 15–30 minutes to ensure complete dissolution.

Detailed protocol and best practices:

• Reconstitution:

  • Add sterile water or PBS directly to the lyophilized IGF-II protein to achieve a concentration of at least 0.1–0.2 mg/mL (100–200 μg/mL).
  • Gently swirl or invert the vial to mix. Do not vortex, as this can denature the protein.
  • Allow the solution to sit at room temperature for 15–30 minutes to ensure the protein is fully dissolved.
  • If the protein is not fully dissolved, gently pipette up and down or swirl again.

• Aliquoting and Storage:

  • Once reconstituted, aliquot the solution into single-use volumes to avoid repeated freeze-thaw cycles, which can degrade the protein.
  • For short-term storage (up to 1 week), keep aliquots at 2–8°C.
  • For long-term storage (up to 3 months), store aliquots at –20°C to –80°C.
  • If further dilution is needed for cell culture, use sterile cell culture medium or PBS containing 0.1% BSA to stabilize the protein and prevent adsorption to plastic.

• Preparation for Cell Culture:

  • Thaw aliquots on ice and dilute to the desired working concentration in cell culture medium immediately before use.
  • Avoid repeated freeze-thaw cycles by using single-use aliquots.
  • If using for sensitive applications, confirm the absence of endotoxin contamination (should be <0.005 EU/μg for most cell culture work).

• Additional Notes:

  • The lyophilized protein may appear as a film or powder; this does not affect quality.
  • Always check the specific product datasheet for any unique requirements, such as reconstitution buffer or additives (e.g., some preparations may recommend 10 mM HCl or inclusion of carrier proteins for stability).
  • For best results, filter-sterilize the reconstituted solution if sterility is critical and the protein is compatible with filtration.

Summary Table:

These guidelines ensure maximum stability and biological activity of recombinant Mouse IGF-II for cell culture applications.