Recombinant Mouse BMPR-IA

Recombinant Mouse BMPR-IA (also known as ALK-3) is a valuable tool for bone morphogenetic protein (BMP) signaling research due to its critical biological roles and well-characterized functions in multiple physiological systems.

Key Research Applications

Developmental and Morphogenetic Studies

Tissue-specific conditional knockout experiments have demonstrated the importance of BMPR-IA in the development and morphogenesis of the heart, lung, palate, and other tissues. This makes recombinant BMPR-IA particularly useful for investigating how BMP signaling regulates developmental processes and organ formation.

Bone Biology Research

BMPR-IA serves as a potent receptor for BMP-2 and plays essential roles in skeletal development, homeostasis, and regeneration. Recombinant BMPR-IA allows you to study BMP signaling at physiological levels and understand the complex interplay between bone formation and resorption. Research has shown that BMP signaling through BMPR-IA is essential for postnatal maintenance of articular cartilage, making it valuable for osteoarthritis and cartilage regeneration studies.

Signaling Pathway Investigation

The protein enables detailed examination of canonical BMP signaling cascades, including Smad phosphorylation and downstream target activation such as Id1 and Sost expression. This is particularly useful for understanding how BMPR-IA-dependent signaling affects osteoblast and osteoclast function through pathways like RANKL-OPG regulation.

Disease Model Studies

Recombinant BMPR-IA is applicable to investigating pathological conditions. For instance, understanding BMPR-IA function is relevant to pulmonary arterial hypertension research, where BMPR-II dysfunction plays a central role in disease pathogenesis.

Bioactivity Validation

The recombinant protein has been validated for bioactivity assays, making it suitable for functional studies examining receptor activation, ligand binding, and downstream signaling responses in cell-based and biochemical assays.

Yes, you can use recombinant mouse BMPR-IA as a standard for quantification or calibration in ELISA assays, provided it is of high purity and its concentration is accurately known. This is a common practice in ELISA development and quantification protocols.

Key considerations and supporting details:

• Recombinant proteins are widely used as ELISA standards: Guidelines for ELISA standard preparation recommend using a purified protein, and if unavailable, a recombinant protein of known concentration is suitable for generating a standard curve. This allows for quantification of the target analyte in unknown samples by comparison to the standard curve.

• BMPR-IA/ALK-3 recombinant proteins are specifically mentioned for ELISA standards: Product information for recombinant mouse BMPR-IA/ALK-3 Fc chimera proteins states that they are suitable for use as ELISA standards, especially when formulated with BSA as a carrier protein. This ensures stability and compatibility with typical ELISA buffers.

• Validation and compatibility: Before using any recombinant protein as a standard, confirm that:

  • The recombinant protein matches the epitope recognized by the capture and detection antibodies in your ELISA.
  • The protein is of high purity (typically >95% by SDS-PAGE).
  • The protein is reconstituted and diluted according to the manufacturer’s instructions to ensure accurate concentration.
  • The standard curve generated with the recombinant protein is linear and covers the expected concentration range of your samples.

• Matrix effects and controls: When quantifying endogenous BMPR-IA in complex samples (e.g., serum or plasma), include spike controls by adding the recombinant standard to the sample matrix to assess recovery and potential matrix effects.

• Interference and specificity: Some ELISA kits report that recombinant BMPR-IA can interfere at high concentrations (>10 ng/mL) in assays designed for related proteins (e.g., BMP-2), so ensure your assay is specific for BMPR-IA and validated for use with the recombinant standard.

Summary of best practices:

• Use a highly purified, well-characterized recombinant BMPR-IA protein.
• Prepare and store the standard according to the supplier’s instructions.
• Validate the standard curve in your specific ELISA setup.
• Include appropriate controls to confirm quantification accuracy.

If your recombinant mouse BMPR-IA meets these criteria, it is appropriate for use as a standard in ELISA quantification and calibration.

Recombinant Mouse BMPR-IA (Bone Morphogenetic Protein Receptor Type IA) has been validated for use in bioactivity assays, cell analysis methods, and as a tool in cell and gene therapy research.

Key validated applications in published research include:

• Bioactivity assays: The recombinant BMPR-IA protein is commonly used to assess ligand-receptor interactions, particularly with BMP-2 and related ligands, to confirm receptor functionality and downstream signaling activation.
• Cell analysis methods: It is utilized in studies involving cell surface receptor recognition, quantification of receptor-ligand binding, and analysis of receptor-mediated cellular responses such as differentiation and gene expression.
• Cell and gene therapy research: BMPR-IA is employed in protocols investigating BMP signaling pathways in stem cells, bone marrow stromal cells, and other progenitor cell types, especially in the context of osteogenesis, adipogenesis, and tissue regeneration.
• ELISA standard: The protein is recommended for use as a standard in ELISA assays to quantify BMPs or their receptors in biological samples.

Supporting details from published research:

• Facilitated receptor recognition: Studies have used recombinant BMPR-IA to investigate how BMP-2 binds and activates its receptor, influencing cellular differentiation and gene expression in cell models such as C2C12 myoblasts.
• Bone marrow stromal cell signaling: Research has validated BMPR-IA for tracking receptor expression and function in aging and disease models, including osteoporosis, using techniques like Western blotting, confocal microscopy, and fluorescent ligand binding.
• Stem cell differentiation: BMPR-IA has been used to dissect its role in adipogenic and osteogenic differentiation of mouse adipose-derived stem cells, often in combination with BMP-2 stimulation and gene knockout approaches.
• ELISA and cell culture: The protein is recommended for use in ELISA assays and cell culture experiments to study BMP signaling and receptor-ligand interactions.

Summary of typical protocols:

• Bioactivity validation: Recombinant BMPR-IA is tested for its ability to bind BMP ligands and activate downstream signaling, often measured by alkaline phosphatase activity, gene expression (qPCR), or immunofluorescence.
• Cell-based assays: Used in cell lines or primary cells to study receptor-mediated effects, including differentiation, proliferation, and apoptosis.
• ELISA standardization: Employed as a reference protein for quantifying BMPs or BMPRs in biological samples.

Note: While recombinant BMPR-IA is validated for these applications, specific experimental protocols and validation data should be referenced from individual studies for precise methodological details.

To reconstitute and prepare Recombinant Mouse BMPR-IA protein (often supplied as a lyophilized Fc chimera) for cell culture experiments, follow these steps:

• Reconstitution:
  Add sterile PBS to the lyophilized protein to achieve a final concentration of 100 μg/mL. Before opening, briefly centrifuge the vial to collect the powder at the bottom.

• Dissolving:
  Gently swirl or invert the vial to dissolve the protein. Avoid vigorous shaking or vortexing, as this can cause foaming and potential denaturation.

• Incubation:
  Allow the solution to sit at room temperature for 15–30 minutes with gentle agitation to ensure complete dissolution.

• Aliquoting and Storage:

  • Prepare aliquots to avoid repeated freeze-thaw cycles.
  • Store aliquots at 2–8 °C for up to 1 month under sterile conditions, or at –20 to –70 °C for up to 3 months.
  • Avoid storing at room temperature or repeated freeze-thawing, as this can reduce protein activity.

• Preparation for Cell Culture:

  • If your protocol requires, dilute the reconstituted stock further in cell culture medium immediately before use.
  • For sensitive applications, consider adding a carrier protein (e.g., 0.1–1% BSA) to minimize adsorption and loss, especially at low working concentrations.
  • Filter the final working solution through a 0.2 μm sterile filter if sterility is critical.

Additional Notes:

• The protein is typically supplied lyophilized from a solution containing PBS and BSA as a carrier.
• Always consult the specific product datasheet for any unique instructions, as formulations can vary slightly between preparations.
• For functional assays, confirm the absence of endotoxin if your cell type is sensitive.

Summary Table: Recombinant Mouse BMPR-IA Reconstitution

These steps will ensure your recombinant BMPR-IA protein is properly prepared for reliable and reproducible cell culture experiments.