Recombinant Mouse IGFBP-2

Recombinant Mouse IGFBP-2 is widely used in research because it is a key regulator of insulin-like growth factor (IGF) signaling, with roles in development, metabolism, cancer biology, and cell signaling. Its use enables precise, controlled studies of IGFBP-2 function in vitro and in vivo.

Key reasons to use recombinant Mouse IGFBP-2 in research applications:

• IGF Modulation: IGFBP-2 binds IGF-I and IGF-II with high affinity, modulating their bioavailability and activity in cell culture and animal models. This allows researchers to dissect IGF-dependent and IGF-independent pathways in processes such as growth, differentiation, and metabolism.
• Developmental and Tissue-Specific Studies: IGFBP-2 is highly expressed in the central nervous system during development and in various tissues in adults, making it relevant for studies of neurobiology, tissue regeneration, and developmental biology.
• Cancer Research: IGFBP-2 has dual roles in cancer, acting as a tumor suppressor in some contexts (e.g., bladder cancer) and as a promoter of tumorigenesis in others (e.g., glioma, pancreatic cancer). Recombinant IGFBP-2 enables functional assays to study its effects on cell proliferation, invasion, and epithelial-to-mesenchymal transition.
• Redox and Post-Translational Regulation: Recombinant IGFBP-2 can be used to study redox-sensitive signaling, as its cysteine residues are subject to redox modifications that affect its function.
• Bioassays and ELISA Standards: Recombinant protein provides a standardized, defined reagent for quantitative bioassays, ELISA calibration, and mechanistic studies.
• Metabolic and Endocrine Research: Overexpression or exogenous addition of IGFBP-2 in animal models has been shown to reduce IGF-I bioavailability, impact glucose homeostasis, and regulate postnatal growth, making it valuable for metabolic studies.
• Complement Pathway and Immune Studies: IGFBP-2 interacts with other IGFBPs to activate immune pathways, such as complement activation in macrophages, relevant for immunology and kidney disease research.

Typical applications include:

• Cell culture experiments to modulate IGF signaling.
• In vivo studies to assess physiological or pathological roles of IGFBP-2.
• Mechanistic studies of cancer progression, metastasis, and chemosensitivity.
• ELISA and other immunoassays as a standard or control.
• Investigation of redox biology and post-translational modifications.

Using recombinant Mouse IGFBP-2 ensures reproducibility, species specificity, and the ability to perform dose-response and mechanistic studies under controlled conditions.

Yes, recombinant mouse IGFBP-2 can be used as a standard for quantification or calibration in ELISA assays, provided it is properly validated for your specific assay system. This is a common practice in commercial mouse IGFBP-2 ELISA kits, where recombinant mouse IGFBP-2 is routinely used to generate standard curves for quantification.

Key considerations and supporting details:

• Standard Curve Preparation: Commercial mouse IGFBP-2 ELISA kits typically use recombinant mouse IGFBP-2 as the standard, with calibration ranges such as 0.031–2 ng/mL or 179.68–11,500 pg/mL. The standards are reconstituted and serially diluted to generate a standard curve, which is then used to interpolate the concentration of IGFBP-2 in unknown samples.

• Assay Validation: It is essential to ensure that the recombinant protein you use as a standard is of high purity, correctly folded, and biologically active, as these factors can affect antibody recognition and quantification accuracy. The recombinant standard should match the native protein as closely as possible in terms of immunoreactivity.

• Matrix Effects: When using recombinant standards, be aware of potential matrix effects—differences in signal due to the sample matrix (e.g., serum, plasma, cell culture supernatant) compared to the standard diluent. Internal controls and spike-recovery experiments are recommended to confirm that the recombinant standard behaves similarly to endogenous IGFBP-2 in your sample matrix.

• Documentation: Always document the source, lot, and concentration of the recombinant standard used, and include a full calibration curve in each assay run for reproducibility and traceability.

• Controls: Include appropriate low and high controls in each assay to ensure the accuracy and precision of quantification.

Summary Table: Use of Recombinant Mouse IGFBP-2 as ELISA Standard

In summary: Using recombinant mouse IGFBP-2 as a standard is scientifically valid and widely accepted for ELISA quantification, provided you validate its performance in your specific assay context and follow best practices for calibration and quality control.

Recombinant Mouse IGFBP-2 has been validated for several key applications in published research, primarily in the context of bioassays, cell culture, in vivo studies, and as a standard in ELISA.

Validated Applications:

• Bioassay:
  Recombinant Mouse IGFBP-2 is frequently used in bioassays to assess its ability to inhibit the biological activity of IGF-I and IGF-II, particularly on cell lines such as MCF-7 human breast cancer cells. This application is central to studies investigating IGFBP-2’s modulation of IGF signaling and its downstream effects on cell proliferation and differentiation.

• Cell Culture:
  IGFBP-2 has been applied in cell culture systems to study its effects on cellular processes, including cytokine secretion from gingival fibroblasts and its impact on osteoclast activity. It is also used to investigate metabolic regulation and insulin sensitivity in adipocytes and other cell types.

• In Vivo Studies:
  Recombinant Mouse IGFBP-2 has been validated in animal models to study its role in growth regulation, metabolic processes, and disease states. For example, it has been used to demonstrate inhibition of GH-stimulated growth in transgenic mice, and to investigate its effects on tissue expansion and regeneration.

• ELISA (Standard):
  Recombinant Mouse IGFBP-2 is used as a standard in ELISA assays for the quantitative detection of IGFBP-2 in mouse and rat serum. This enables precise measurement of IGFBP-2 levels in various experimental contexts.

Additional Context:

• Mechanistic Studies:
  IGFBP-2 is utilized to dissect molecular pathways, such as its interaction with IGF, modulation of insulin sensitivity, and involvement in complement activation in macrophages. It is also used to study its binding to glycosaminoglycans and cell surface receptors in developmental and oncogenic signaling.

• Disease Models:
  Applications include research into obesity, insulin resistance, metabolic syndrome, diabetes, and cancer, where IGFBP-2’s regulatory role is explored both in vitro and in vivo.

Summary Table of Validated Applications

These applications are supported by multiple peer-reviewed studies and are widely adopted in research investigating IGFBP-2’s biological functions and disease relevance.

To reconstitute and prepare Recombinant Mouse IGFBP-2 protein for cell culture experiments, follow these best practices based on manufacturer guidelines and scientific protocols:

Reconstitution:

1. Centrifuge the vial briefly before opening to ensure all lyophilized powder is at the bottom.
2. Reconstitute the protein in sterile distilled water or sterile PBS (pH 7.4), depending on the formulation. Most protocols recommend a final concentration of 0.1–0.5 mg/mL.
   • For some products, reconstitution at 100 µg/mL in sterile PBS is also acceptable.
3. Gently mix the solution by pipetting up and down or by inverting the vial. Do not vortex or pipette vigorously to avoid denaturation or foaming.

Preparation for Cell Culture:

• If the protein will be used for sensitive cell culture applications, consider adding a carrier protein (e.g., 0.1% BSA, 5% HSA, or 5% trehalose) to stabilize the solution and minimize adsorption losses.
• Filter sterilize the reconstituted protein solution using a 0.22 µm filter if not already sterile.
• Aliquot the solution to avoid repeated freeze-thaw cycles, which can degrade the protein.

Storage:

• Store reconstituted protein at 2–8°C for up to one month if used soon.
• For longer storage, keep aliquots at –20°C to –70°C in a manual defrost freezer.
• Avoid repeated freeze-thaw cycles.

Usage in Cell Culture:

• Dilute the reconstituted protein in cell culture medium to the desired working concentration.
• Ensure compatibility with your cell type and experimental conditions.

Additional Notes:

• Always refer to the specific product datasheet for any unique requirements.
• If using for functional assays (e.g., IGF inhibition), validate activity with a biological assay.

By following these steps, you will ensure optimal activity and stability of Recombinant Mouse IGFBP-2 for your cell culture experiments.