Recombinant Human PDGF Rα

Recombinant Human PDGF Rα (Platelet-Derived Growth Factor Receptor Alpha) is a valuable tool in research because it enables precise investigation of PDGF signaling pathways, receptor-ligand interactions, and downstream cellular effects relevant to tissue repair, fibrosis, oncology, and developmental biology.

Key reasons to use recombinant human PDGF Rα in research applications:

• Mechanistic Studies of PDGF Signaling: PDGF Rα is a tyrosine kinase receptor that mediates cellular responses to PDGF ligands, including proliferation, migration, and survival in various cell types such as hepatic stellate cells, fibroblasts, and mesenchymal cells. Using recombinant PDGF Rα allows you to dissect the specific contributions of this receptor in signaling cascades, independent of endogenous receptor expression.

• Receptor-Ligand Binding Assays: Soluble recombinant PDGF Rα can be used to study binding kinetics and specificity with different PDGF isoforms (e.g., PDGF-AA, PDGF-BB), enabling quantitative analysis of ligand-receptor interactions and competitive inhibition studies.

• Decoy Receptor Applications: The soluble form of recombinant PDGF Rα can act as a decoy receptor, sequestering PDGF ligands and preventing them from activating cell-surface receptors. This approach is useful for investigating the biological consequences of PDGF pathway inhibition in vitro and in vivo, such as reduced mitogenesis, migration, or fibrotic responses.

• Modeling Disease Mechanisms: PDGF Rα is implicated in pathological processes such as fibrosis, cancer progression, and tissue remodeling. Recombinant PDGF Rα enables controlled experiments to model these mechanisms, test inhibitors, or evaluate therapeutic strategies targeting PDGF signaling.

• Cellular and Molecular Assays: Recombinant PDGF Rα can be used in ELISA, Western blot, immunoprecipitation, and cell-based assays to quantify receptor activation, downstream phosphorylation events, and gene expression changes in response to PDGF stimulation.

• Therapeutic Development: Understanding the role of PDGF Rα in disease models supports the development of targeted therapies, such as monoclonal antibodies or small molecules that modulate PDGF Rα activity.

Summary of Applications:

• Dissecting PDGF signaling pathways
• Quantifying ligand-receptor interactions
• Acting as a decoy receptor in functional assays
• Modeling fibrotic, oncogenic, or regenerative processes
• Screening and validating PDGF pathway inhibitors

Scientific Context:
PDGF Rα is upregulated in various pathological conditions (e.g., chronic liver injury, cardiac repair, cancer), and its activation triggers multiple downstream pathways (Erk1/2, Akt, mTOR, FAK, etc.) that regulate cell proliferation, migration, and tissue remodeling. Recombinant forms provide a controlled, reproducible reagent for mechanistic and translational research.

In summary, recombinant human PDGF Rα is essential for precise, mechanistic, and translational studies of PDGF-driven biology and pathology, supporting both basic research and therapeutic innovation.

Yes, recombinant human PDGF Rα can be used 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 or recombinant protein to generate the standard curve, as this allows for accurate quantification of the target analyte.
• Product formulations for ELISA: Recombinant human PDGF Rα is often supplied in formulations specifically recommended for use as ELISA standards. For example, a commonly used formulation includes BSA as a carrier protein, which is advised for ELISA applications to enhance stability and reproducibility.
• Standard curve generation: When using recombinant PDGF Rα as a standard, prepare a dilution series according to your assay’s protocol to generate a standard curve. This curve is then used to interpolate the concentration of PDGF Rα in your samples.
• Validation and parallelism: It is important to confirm that your samples show parallelism with the standard curve, ensuring that the recombinant standard behaves similarly to the native protein in your assay matrix. This is typically assessed by running serial dilutions of both the standard and representative samples.
• Concentration range: ELISA kits for PDGF Rα typically use standard curves in the range of approximately 0.5–120 ng/mL, but this can be adjusted based on your expected sample concentrations and assay sensitivity.
• Carrier-free vs. carrier-added: If your assay is sensitive to carrier proteins (such as BSA), use a carrier-free recombinant standard. Otherwise, the BSA-containing formulation is generally suitable and may improve protein stability.

Best practices:

• Reconstitute the recombinant standard exactly as instructed by the supplier to ensure accurate concentration.
• Validate the standard curve’s linearity and dynamic range for your specific assay conditions.
• Confirm that the recombinant standard is recognized equivalently by the antibodies used in your ELISA.

In summary: Recombinant human PDGF Rα is suitable and commonly used as a standard for ELISA quantification, provided it is appropriately formulated and validated for your assay system.

Recombinant Human PDGF Rα has been validated for several key applications in published research, primarily in bioassays, cell culture, ELISA standards, and kinase assays.

Validated Applications in Published Research:

• Bioassays:
  Used to study ligand-receptor interactions, signal transduction, and cellular responses such as proliferation, migration, and differentiation in various cell types, including fibroblasts, mesenchymal stromal cells, and melanoma cells. For example, PDGF Rα has been used to assess phosphorylation of downstream signaling molecules (Akt, PKC) in hepatic stellate cells and to investigate antiproliferative effects in melanoma cell lines.

• Cell Culture:
  Applied as a functional reagent to stimulate or inhibit cellular processes in vitro, including maintenance and differentiation of stem/immune cells, and to model disease states such as neurodegeneration and wound healing. It is also used to differentiate human iPS cells into oligodendrocyte precursors and to study mesenchymal stem cell heterogeneity.

• ELISA Standard:
  Recombinant PDGF Rα is recommended as a standard for quantifying PDGF receptor levels in immunoassays, enabling precise measurement of receptor expression in biological samples.

• Kinase Assays:
  Utilized to measure receptor tyrosine kinase activity, including phosphorylation events and downstream signaling, which are critical for understanding receptor function and drug screening.

Additional Context:

• Functional Studies:
  Recombinant PDGF Rα has been used to dissect receptor-ligand specificity, showing preferential binding to PDGF-AA and PDGF-CC, and to study the effects of receptor overexpression or inhibition in cancer and tissue regeneration models.

• Disease Modeling:
  Applications include investigating the role of PDGF Rα in chronic liver injury, cancer cell proliferation, angiogenesis, and wound healing, both in vitro and in vivo.

• Protein Characterization:
  Recombinant PDGF Rα is suitable for Western blot analysis to confirm protein expression and integrity.

Summary Table:

These applications are supported by multiple peer-reviewed studies and technical datasheets, confirming the versatility of recombinant human PDGF Rα in both basic and translational research.

To reconstitute and prepare Recombinant Human PDGF Rα protein for cell culture experiments, follow these best-practice steps based on current protocols and manufacturer recommendations:

1. Centrifuge the vial briefly before opening to ensure all lyophilized protein is at the bottom.

2. Reconstitution buffer:

   • Most protocols recommend sterile phosphate-buffered saline (PBS) as the reconstitution buffer for PDGF Rα.
   • If the datasheet specifies, use the buffer indicated in the Certificate of Analysis (CoA) for your specific product.
   • For some applications, a small amount of carrier protein such as 0.1% bovine serum albumin (BSA) or human serum albumin (HSA) in PBS may be added to stabilize the protein, especially for further dilutions.

3. Reconstitution concentration:

   • A typical reconstitution concentration is 100 μg/mL in sterile PBS.
   • For example, to reconstitute 50 μg of protein, add 0.5 mL of PBS to achieve 100 μg/mL.

4. Dissolving the protein:

   • Gently pipette the buffer down the side of the vial to avoid foaming.
   • Allow the vial to sit at room temperature for 15–30 minutes with gentle agitation (do not vortex).
   • Ensure the protein is fully dissolved before proceeding.

5. Aliquoting and storage:

   • Divide the reconstituted protein into small aliquots to avoid repeated freeze-thaw cycles.
   • Store aliquots at –20°C to –80°C for long-term use.
   • Avoid multiple freeze-thaw cycles, as this can reduce protein activity.

6. Working solution preparation:

   • For cell culture, dilute the stock solution to the desired working concentration using cell culture medium or PBS containing 0.1% BSA/HSA to prevent adsorption to plasticware.
   • Filter sterilize the final working solution if sterility is required.

Summary Table: Key Steps for PDGF Rα Reconstitution

Important notes:

• Always consult the product-specific datasheet or CoA for any unique requirements.
• If using for functional cell assays, confirm that the buffer and additives are compatible with your cell type and assay conditions.

If you need a protocol for a specific application (e.g., ELISA standard, cell signaling), the buffer and concentration may differ slightly. Always adjust according to your experimental needs and the guidance provided with your recombinant protein.