Recombinant Mouse Noggin

Recombinant Mouse Noggin is widely used in research applications because it is a potent and specific antagonist of bone morphogenetic proteins (BMPs), particularly BMP-2, BMP-4, BMP-7, and others, which are critical regulators of cell fate, differentiation, and tissue development.

Key scientific reasons to use recombinant mouse Noggin:

• BMP Pathway Inhibition: Noggin binds BMPs with high affinity, preventing them from interacting with their receptors and thereby inhibiting BMP signaling. This is essential for controlling differentiation pathways in stem cell cultures, especially when maintaining pluripotency or directing specific lineage commitment.

• Stem Cell Culture and Differentiation: Mouse Noggin is crucial for the culture of mouse and rat embryonic stem cells, ensuring reproducible and physiologically relevant cell cultures. It supports the maintenance of undifferentiated stem cells and can promote differentiation into neural, retinal, endodermal, adipogenic, and cardiomyocyte lineages, depending on the protocol and context.

• Developmental Biology Studies: Noggin is indispensable for studying embryonic development, as it regulates neural tube formation, somite patterning, and the development of ectodermal and mesodermal structures such as neural tissue, teeth, hair follicles, and skeletal elements.

• Tissue Engineering and Regeneration: By modulating BMP activity, Noggin can influence chondrogenesis, osteogenesis, and joint formation, making it valuable in skeletal regenerative therapies and cartilage/bone tissue engineering.

• Disease Modeling and Functional Assays: Noggin is used in bioassays to study BMP-related signaling in disease models, including osteoarthritis, neural injury, and cancer, as well as in functional assays for cell migration, proliferation, and differentiation.

• Species-Specific Activity: Recombinant mouse Noggin is optimized for mouse and rat systems, ensuring species-specific activity and reproducibility in experimental protocols involving these models.

Additional considerations:

• High Purity and Defined Activity: Recombinant preparations offer high purity and consistent bioactivity, which is critical for reproducible results in sensitive cell culture and differentiation protocols.
• Animal Component-Free Options: Some recombinant mouse Noggin products are available in animal component-free formulations, reducing variability and risk of contamination in defined culture systems.

Summary of applications:

• Maintenance and differentiation of embryonic and induced pluripotent stem cells.
• Neural, retinal, endodermal, adipogenic, and cardiomyocyte differentiation protocols.
• Developmental biology and tissue patterning studies.
• Skeletal and cartilage tissue engineering.
• Disease modeling and functional bioassays.

Using recombinant mouse Noggin enables precise control over BMP signaling, which is fundamental for many cell biology, developmental, and regenerative research applications.

Yes, you can use recombinant Mouse Noggin as a standard for quantification or calibration in ELISA assays, provided that the recombinant protein is suitable for this purpose and is compatible with your specific ELISA kit.

Key Points:

• Recombinant Mouse Noggin is commonly used as a standard in ELISA assays to generate a standard curve for quantifying endogenous Noggin in samples such as serum, plasma, or cell culture supernatants.
• Many commercial ELISA kits for Mouse Noggin (e.g., Invitrogen, Assay Genie, RayBiotech, Cusabio) are designed to detect both natural and recombinant forms of the protein, making recombinant Noggin appropriate for use as a standard.
• For best results, use a recombinant Mouse Noggin protein that is lyophilized and formulated with a carrier protein (such as BSA), as this formulation is recommended for use in cell culture and as an ELISA standard. Carrier-free versions are typically reserved for applications where BSA could interfere.
• Always check the datasheet or product specifications of both your recombinant protein and your ELISA kit to confirm compatibility and recommended dilution ranges.

Example:

• Bio-Techne/R&D Systems Recombinant Mouse Noggin (1967-NG): Formulated with BSA, suitable for use as an ELISA standard.
• BioLegend Recombinant Mouse Noggin (carrier-free): Not recommended for ELISA standard use unless specifically validated for this purpose.

Best Practice:

• Prepare a dilution series of the recombinant Noggin standard according to the assay range of your ELISA kit (e.g., 0.23–50 ng/mL).
• Run the standard curve alongside your samples in each assay to ensure accurate quantification.

In summary, yes, recombinant Mouse Noggin can be used as a standard for ELISA quantification, but ensure it is the appropriate formulation and compatible with your assay system.

Recombinant Mouse Noggin has been validated for a wide range of applications in published research, primarily focusing on cell culture, differentiation, and functional assays. Key applications include:

• Neural stem cell and retinal cell differentiation from ES/iPS cells.
• Endodermal progenitor differentiation.
• Adipogenic differentiation of mesenchymal stem cells (MSCs).
• Cardiomyocyte differentiation of mouse embryonic stem (ES) cells.
• Bioassays to study BMP signaling inhibition and its effects on cell behavior.
• Cell migration and motility studies.
• Cell proliferation assays.
• Stem and immune cell functional studies.
• Neuroprotective and neurorestorative therapy research, including studies on ischemic stroke and microglial activation.
• Cartilage and joint development studies, including inhibition of chondrocyte differentiation.
• Tissue repair and remodeling in models of brain injury and osteoarthritis.

These applications demonstrate the versatility of Recombinant Mouse Noggin in developmental biology, regenerative medicine, and disease modeling research.

To reconstitute and prepare Recombinant Mouse Noggin protein for cell culture experiments, dissolve the lyophilized protein at 100 μg/mL in sterile PBS containing at least 0.1% BSA or HSA as a carrier protein. This carrier protein helps stabilize Noggin and prevents adsorption to plasticware, which is critical for bioactivity in cell culture applications.

Step-by-step protocol:

• Centrifuge the vial briefly before opening to ensure all powder is at the bottom.
• Warm the vial to room temperature before opening to minimize condensation.
• Add sterile PBS (pH 7.2–7.4) containing 0.1–1% BSA or HSA to achieve a final concentration of 100 μg/mL.
• Gently mix by pipetting up and down or gentle inversion. Do not vortex or shake vigorously, as this may denature the protein.
• Allow the solution to sit at room temperature for several minutes to ensure complete dissolution. If solubility issues persist, incubate at 4°C overnight.
• Aliquot the reconstituted protein into working volumes to avoid repeated freeze-thaw cycles.
• Store aliquots at –20°C or below. For short-term use (up to 1 week), storage at 2–8°C is acceptable.

Additional notes:

• If your Noggin preparation is carrier-free (no BSA/HSA), you may reconstitute in sterile PBS alone, but adding carrier protein is recommended for cell culture to maintain stability and activity.
• For some preparations, reconstitution in 10 mM acetic acid or sterile distilled water is also acceptable, but PBS with carrier protein is preferred for cell culture.
• Do not exceed 1 mg/mL protein concentration during reconstitution to avoid solubility issues.
• Confirm protein integrity by SDS-PAGE if needed.

Working concentrations for cell culture:
Typical effective concentrations for Noggin in cell culture range from 0.04–0.16 μg/mL for BMP inhibition assays, but optimal dosing should be determined empirically for your specific application.

Summary Table:

This protocol ensures maximum stability and bioactivity of recombinant Mouse Noggin for cell culture experiments.