Recombinant Human PDGF Rβ

Recombinant Human PDGF Rβ (Platelet-Derived Growth Factor Receptor Beta) is used in research to study and manipulate key cellular processes such as proliferation, migration, survival, and tissue regeneration, particularly in contexts involving mesenchymal cells, vascular biology, immunology, and tissue engineering.

Key scientific applications and rationale:

• Cellular Signaling and Mechanistic Studies: PDGF Rβ is a critical receptor mediating the effects of PDGF ligands (such as PDGF-BB and PDGF-DD), which are potent mitogens and chemoattractants for mesenchymal cells, including vascular smooth muscle cells and fibroblasts. Using recombinant PDGF Rβ allows precise dissection of downstream signaling pathways (e.g., ERK, AKT) and their roles in cell proliferation, migration, and survival.

• Tissue Regeneration and Repair: Recombinant human PDGF (and by extension, its receptor PDGF Rβ) has demonstrated efficacy in promoting tissue regeneration, including bone repair, skin wound healing, and periodontal tissue regeneration. The receptor is essential for mediating these effects, making it a valuable tool for studying regenerative mechanisms and developing new therapeutic strategies.

• Immunology and Cell Survival: Recent research shows that PDGF-D–PDGFRβ signaling enhances the survival of human natural killer (NK) cells in response to IL-15, without affecting their effector functions. This has implications for improving NK cell-based immunotherapies and understanding immune cell homeostasis.

• Vascular Biology and Angiogenesis: PDGF Rβ is predominantly expressed on vascular smooth muscle cells and pericytes, where it regulates vascular development, stability, and remodeling. Recombinant PDGF Rβ can be used to model vascular diseases, study angiogenesis, and test anti-angiogenic compounds.

• Disease Modeling: Aberrant PDGF/PDGFRβ signaling is implicated in pathological conditions such as atherosclerosis, fibrosis, cancer, and proliferative retinopathies. Recombinant PDGF Rβ enables in vitro and in vivo modeling of these diseases, facilitating drug screening and mechanistic studies.

Best practices and considerations:

• Use recombinant PDGF Rβ in controlled in vitro systems to study ligand-receptor interactions, downstream signaling, and cellular responses.
• Combine with specific PDGF ligands (e.g., PDGF-BB, PDGF-DD) to dissect isoform-specific effects and receptor activation dynamics.
• Employ in co-culture or tissue engineering models to investigate cell–cell interactions and tissue-level responses.

Summary:
Using recombinant human PDGF Rβ in research provides a powerful tool to elucidate the molecular mechanisms of cell growth, migration, survival, and tissue regeneration, with broad applications in regenerative medicine, immunology, vascular biology, and disease modeling.

You can use recombinant human PDGF Rβ as a standard for quantification or calibration in your ELISA assays, provided it is highly purified and its concentration is accurately determined. This is a common and accepted practice in quantitative ELISA, where a standard curve is generated using known concentrations of a purified or recombinant protein to enable precise quantification of the target analyte in your samples.

Key considerations:

• Purity and Source: The recombinant PDGF Rβ should be of high purity, ideally similar to the protein detected in your samples, and its concentration must be reliably measured (e.g., by absorbance at 280 nm, BCA, or Bradford assay).
• Standard Curve Preparation: Prepare a serial dilution of the recombinant protein to generate a standard curve covering the expected range of your sample concentrations.
• Matrix Effects: If your samples are in a complex matrix (e.g., serum, plasma, tissue lysate), consider preparing your standards in the same matrix or include matrix-matched controls to account for potential interference.
• Validation: Confirm that the recombinant standard behaves similarly to the endogenous protein in your assay system. This includes comparable binding to capture/detection antibodies and similar signal generation.
• Documentation: Many commercial ELISA kits for PDGF Rβ are calibrated using highly purified recombinant human PDGF Rβ, demonstrating the suitability of this approach.

Limitations:

• If your recombinant PDGF Rβ is not fully glycosylated or structurally identical to the native protein (e.g., expressed in E. coli vs. mammalian cells), minor differences in antibody recognition or assay performance may occur. This is generally not a problem for most quantitative ELISAs but should be validated for your specific application.

Summary:
Using recombinant human PDGF Rβ as a standard is scientifically valid and widely practiced for ELISA quantification, as long as you ensure its purity, accurate quantification, and comparable immunoreactivity to the native protein in your assay.

Recombinant Human PDGF Rβ (Platelet-Derived Growth Factor Receptor Beta) has been validated in published research for several key applications, primarily in cell signaling studies, quantification of receptor expression, and functional assays related to cell proliferation, migration, and survival.

Key validated applications include:

• Quantitative RT-PCR and ELISA: Recombinant human PDGF Rβ has been used as a standard or control in quantitative RT-PCR and ELISA assays to measure mRNA and protein expression levels of PDGF receptors in human cells, particularly in studies of vascular smooth muscle cells and atherosclerosis.

• Cell Signaling and Functional Assays: It is widely used to study PDGF-BB:PDGFRβ signaling pathways, including downstream effects on cell proliferation, migration, and differentiation. For example, research has validated its use in dissecting the role of PDGFRβ signaling in mesenchymal stem cell (MSC) proliferation and differentiation, as well as in neural and vascular cell models.

• Flow Cytometry and Immunoblotting: Recombinant PDGF Rβ has been used to validate antibody specificity and receptor expression by flow cytometry and immunoblotting, particularly in immune cell studies such as those investigating NK cell survival and function.

• In Vivo and In Vitro Functional Studies: It has been employed to explore the effects of PDGF-D−PDGFRβ signaling on immune cell survival, especially in the context of IL-15–mediated NK cell survival, both in vitro and in animal models.

• Tumor Microenvironment and Cancer Research: PDGFRβ signaling has been studied in the context of tumor stroma, where recombinant PDGF Rβ is used to investigate its role in cancer-associated fibroblast function and tumor invasiveness.

• Neuroscience and Vascular Biology: Studies have validated its use in characterizing PDGF-BB:PDGFRβ signaling in human brain tissue, particularly in neurodegenerative disease models.

Summary Table of Validated Applications

Additional Notes:

• Recombinant PDGF Rβ is often used in combination with its ligands (e.g., PDGF-BB, PDGF-DD) to study receptor-ligand interactions and downstream signaling.
• It is a critical tool for dissecting the molecular mechanisms of tissue regeneration, immune cell regulation, and cancer biology.

If you require protocols or more specific assay details, please specify the experimental context.

To reconstitute and prepare Recombinant Human PDGF Rβ protein for cell culture experiments, first confirm the formulation and recommended diluent from the product datasheet. For most lyophilized PDGF Rβ proteins, reconstitution in sterile PBS at 500 μg/mL is standard for cell culture applications. Carrier-free formulations should be handled with care to avoid protein loss and instability.

Step-by-step protocol:

1. Centrifuge the vial briefly to collect all lyophilized material at the bottom before opening.
2. Add sterile PBS (phosphate-buffered saline) to achieve a final concentration of 500 μg/mL. If the datasheet specifies a different buffer or concentration, follow those instructions.
3. Gently mix by pipetting or slow vortexing. Avoid vigorous shaking to prevent foaming and protein denaturation.
4. Allow the protein to dissolve at room temperature for 15–30 minutes with gentle agitation.
5. Aliquot the reconstituted protein into small working volumes (≥10 μL) to minimize freeze-thaw cycles.
6. Store aliquots at –20°C to –80°C for long-term use. Avoid repeated freeze-thaw cycles, as these can reduce protein activity.
7. For cell culture, dilute the stock solution in cell culture medium immediately before use. If using a carrier-free protein, consider adding 0.1–1% BSA to the medium to stabilize the protein and prevent adsorption to plasticware.

Additional notes:

• Always consult the specific product datasheet for any unique requirements regarding buffer composition, concentration, or additives.
• If the protein is not carrier-free, BSA may already be present to enhance stability.
• For functional assays, ensure the final working concentration matches the requirements of your experimental design.

Summary of key points:

• Reconstitute in sterile PBS at 500 μg/mL (unless otherwise specified).
• Aliquot and store at –20°C to –80°C.
• Minimize freeze-thaw cycles.
• Dilute in cell culture medium with optional BSA for stability.

These steps will help maintain the integrity and biological activity of recombinant PDGF Rβ for cell culture experiments.