Recombinant Mouse PDGF Rα

Using Recombinant Mouse PDGF Rα (Platelet-Derived Growth Factor Receptor alpha) in research applications is valuable for dissecting the roles of PDGF signaling in development, disease, and cellular processes, particularly in mouse models. This recombinant protein enables precise, controlled studies of receptor-ligand interactions, signaling pathways, and functional outcomes in vitro and in vivo.

Key scientific applications and rationale:

• Modeling PDGF Signaling: Recombinant PDGF Rα allows you to study the specific effects of PDGF ligands (such as PDGF-AA and PDGF-BB) on mouse cells by providing a defined receptor population. This is crucial for understanding how PDGF signaling regulates cell proliferation, migration, differentiation, and survival, especially in mesenchymal and neural crest-derived tissues.

• Functional Studies: Overexpression or exogenous addition of PDGF Rα can be used to modulate cellular responses. For example, increased PDGF Rα expression or activation has been shown to induce strong antiproliferative effects, apoptosis, and cell cycle arrest in certain cancer cell lines, such as melanoma, both in vitro and in vivo. This makes it a useful tool for investigating tumor biology and potential therapeutic mechanisms.

• Decoy Receptor/Competitive Inhibition: Soluble recombinant PDGF Rα can act as a decoy receptor, binding PDGF ligands and preventing them from activating cell-surface PDGF receptors. This approach is used to inhibit PDGF-driven signaling pathways, which is valuable in studies of fibrosis, cancer, and tissue regeneration.

• Developmental Biology: PDGF Rα is essential for normal development of multiple organs, including the heart, neural crest, lung, and skeleton. Recombinant forms can be used to probe developmental processes, rescue genetic deficiencies, or study receptor-ligand specificity in mouse embryonic models.

• Fibrosis and Tissue Repair: PDGF Rα signaling is implicated in fibrotic responses and tissue regeneration. Recombinant receptor can be used to modulate or block these pathways in models of liver, cardiac, or other tissue injuries, helping to elucidate mechanisms of fibrosis and repair.

Best practices:

• Use recombinant mouse PDGF Rα when working with mouse cells or tissues to ensure species compatibility and physiological relevance.
• Employ the recombinant receptor in both gain-of-function (overexpression, supplementation) and loss-of-function (decoy, competitive inhibition) experimental designs, depending on your research question.
• Validate biological activity and specificity in your system, as receptor function can vary depending on cell type and context.

Summary of advantages:

• Enables controlled, reproducible studies of PDGF signaling in mouse systems.
• Facilitates mechanistic dissection of receptor-mediated effects in development, disease, and regeneration.
• Provides a tool for both activating and inhibiting PDGF pathways, depending on experimental needs.

These features make recombinant mouse PDGF Rα a versatile and powerful reagent for a wide range of biomedical research applications involving PDGF signaling.

You can use recombinant Mouse PDGF Rα as a standard for quantification or calibration in your ELISA assays, provided that the recombinant protein is of high purity, its concentration is accurately determined, and it is compatible with your assay’s antibodies and detection system.

Key considerations:

• Purity and Quantification: The recombinant protein should be highly purified and its concentration precisely measured, ideally by absorbance at 280 nm or amino acid analysis. Impurities or inaccurate quantification can lead to errors in your standard curve.
• Compatibility: The recombinant PDGF Rα must be recognized by the capture and detection antibodies used in your ELISA. Most commercial ELISA kits are validated with specific recombinant standards; using a different recombinant protein may affect assay performance if epitopes differ or if the protein is not in the correct conformation.
• Formulation: Recombinant proteins are often supplied with or without carrier proteins (such as BSA). For ELISA standards, carrier-free formulations are preferred to avoid interference.
• Validation: If you are substituting the standard provided with a kit, you should validate that your recombinant PDGF Rα generates a standard curve with similar sensitivity, linearity, and parallelism to the kit standard. This ensures accurate quantification of unknowns.

Best practices:

• Prepare serial dilutions of the recombinant protein in the same buffer as your samples and kit standards.
• Confirm that the standard curve covers the expected concentration range of your samples.
• Validate the standard curve for linearity and parallelism with endogenous samples if possible.

Limitations:

• If your recombinant PDGF Rα is not validated for use as an ELISA standard, results may not be directly comparable to those obtained with the kit’s standard, especially if there are differences in glycosylation, folding, or tag sequences.
• Always consult the ELISA kit protocol and, if possible, contact the kit manufacturer for guidance on using alternative standards.

In summary, recombinant Mouse PDGF Rα can be used as an ELISA standard if it is pure, accurately quantified, and validated for compatibility with your assay system. Validation experiments are essential to ensure reliable quantification.

Recombinant Mouse PDGF Rα has been validated in published research for several key applications, primarily in studies of cell signaling, neural stem cell biology, tissue regeneration, and immunodetection assays.

Validated Applications in Published Research:

• Immunohistochemistry (IHC) and Immunofluorescence (IF):

  • Recombinant Mouse PDGF Rα has been used as a target antigen for antibody validation in immunohistochemistry and immunofluorescence, particularly to identify and distinguish PDGFRα-expressing cells in mouse tissues, such as neural stem cells and oligodendrocyte precursor cells in the brain.
  • Studies have used anti-PDGFRα antibodies to stain tissue sections and confirm the presence and distribution of PDGFRα+ cells, validating the specificity of detection using recombinant protein as a control.

• Cell Stimulation and Functional Assays:

  • Recombinant PDGF ligands (such as PDGF-AA and PDGF-BB) are commonly used to stimulate cells expressing PDGF Rα in vitro, to study downstream signaling pathways, cell proliferation, and differentiation responses.
  • For example, recombinant PDGF-AA has been shown to promote proliferation of mouse 3T3 fibroblasts, indicating the functional activity of the receptor in cell-based assays.

• Isolation and Characterization of PDGFRα+ Cells:

  • Protocols for the purification and immunopanning of PDGFRα+ oligodendrocyte precursor cells from mouse brain utilize recombinant PDGF Rα for antibody validation and as a positive control in flow cytometry and cell sorting applications.

• Tissue Regeneration and Wound Healing Models:

  • Recombinant PDGF (including forms that bind PDGF Rα) has been validated in animal models and clinical studies for promoting tissue regeneration, wound healing, and bone repair.
  • These studies demonstrate the biological relevance of PDGF Rα signaling in vivo, with recombinant proteins used to activate the receptor and assess functional outcomes in tissue repair.

• ELISA and Western Blot Standards:

  • Recombinant Mouse PDGF Rα is used as a standard or positive control in ELISA and Western blot assays to quantify or detect PDGFRα protein levels in biological samples.

Summary Table of Validated Applications

Additional Notes:

• The specific recombinant protein format (e.g., full-length, Fc chimera, carrier-free) may influence its suitability for certain applications, such as cell culture or as an ELISA standard.
• While PDGF ligands (AA, BB, CC) are often used to activate PDGF Rα in functional studies, the recombinant receptor itself is primarily used for antibody validation, assay development, and as a standard.

If you require details on a specific application or protocol, please specify the context (e.g., in vitro signaling, in vivo regeneration, antibody validation).

To reconstitute and prepare Recombinant Mouse PDGF Rα protein for cell culture experiments, first confirm the specific formulation and recommended diluent from the product datasheet. Most recombinant PDGF Rα proteins are supplied lyophilized and are typically reconstituted in sterile PBS or sterile distilled water, depending on the manufacturer’s instructions.

General Protocol for Reconstitution and Preparation:

1. Centrifuge the vial briefly to collect all lyophilized powder at the bottom before opening.
2. Add sterile PBS (or the recommended buffer) to the vial. Common reconstitution concentrations are:
   • 100 μg/mL in sterile PBS
   • 500 μg/mL in PBS for some preparations
   • 0.1–1.0 mg/mL in sterile distilled water or PBS is generally suitable for most recombinant proteins
3. Gently mix by pipetting up and down or swirling. Avoid vigorous shaking or vortexing, as this can denature the protein.
4. Allow the protein to dissolve at room temperature for 15–30 minutes with gentle agitation.
5. Aliquot the solution to avoid repeated freeze-thaw cycles, which can degrade the protein.
6. Storage after reconstitution:
   • Store at 2–8 °C for up to 1 month under sterile conditions.
   • For longer-term storage, freeze aliquots at –20 °C to –70 °C for up to 3 months.

Preparation for Cell Culture:

• Before adding to cell cultures, dilute the reconstituted stock to the desired working concentration using sterile cell culture medium or PBS.
• If the protein is to be used in functional assays, consider adding a carrier protein (e.g., 0.1% BSA) to minimize adsorption to plastic and improve stability, unless the application requires carrier-free conditions.
• Filter-sterilize the final working solution if sterility is a concern and the protein is compatible with filtration.

Critical Notes:

• Always consult the specific product datasheet for any unique requirements or recommended buffers, as some formulations may contain stabilizers (e.g., trehalose) or require specific reconstitution conditions.
• Avoid repeated freeze-thaw cycles by aliquoting immediately after reconstitution.
• Do not vortex the protein solution.

Summary Table:

If your application is highly sensitive or requires carrier-free protein, ensure all solutions and consumables are endotoxin-free and protein adsorption is minimized. Always verify the activity of the reconstituted protein in a pilot experiment before large-scale use.