Recombinant Mouse PDGF R-β

Recombinant mouse PDGF receptor-β (PDGFR-β) is a valuable tool for investigating critical cellular signaling pathways and physiological processes. Here are the key reasons to incorporate this protein into your research:

Signal Transduction Studies

PDGFR-β serves as an essential component for studying growth factor signaling mechanisms. The receptor mediates diverse cellular responses including proliferation, migration, and survival through activation of downstream effector kinases such as Akt, ERK1/2, and Src. By using recombinant PDGFR-β, you can examine receptor activation, phosphorylation patterns, and the temporal dynamics of signaling cascade initiation in controlled experimental conditions.

Trafficking and Subcellular Localization Research

This protein is particularly valuable for investigating endocytic trafficking pathways. PDGFR-β undergoes ligand-induced internalization and trafficking to perinuclear late endosomal compartments, where it continues to engage in signaling. Recombinant PDGFR-β enables you to track receptor movement through different cellular compartments and study how trafficking regulates signaling outcomes, which is critical for understanding both normal physiology and disease mechanisms.

Tissue-Specific Applications

PDGFR-β plays specialized roles in multiple biological contexts. The receptor is essential for vascular smooth muscle cell proliferation in response to PDGF stimulation, contributes to blood-brain barrier integrity and functional recovery after cerebral ischemia, and supports natural killer cell survival through IL-15-mediated signaling. This versatility makes recombinant PDGFR-β applicable across diverse research areas including vascular biology, neurovascular disease, and immunology.

Protein-Protein Interaction Analysis

Recombinant PDGFR-β can be used to study receptor dimerization and ligand binding interactions. The protein's ability to form homo- or heterodimerization complexes with PDGF receptor-α makes it suitable for investigating how receptor architecture influences signaling specificity and cellular outcomes.

Mechanistic Disease Research

Understanding PDGFR-β function provides insights into pathological conditions. Research demonstrates that manipulating PDGFR-β signaling affects metabolic function, immune responses, and tissue repair processes, making this protein relevant for investigating disease mechanisms and potential therapeutic interventions.

Recombinant Mouse PDGF R-β protein can be used as a standard for quantification or calibration in ELISA assays, provided it is appropriately validated and formulated for this purpose. Several sources indicate that recombinant proteins are commonly used as standards in ELISA, but there are important technical considerations to ensure accurate quantification.

Key considerations:

• Formulation: Recombinant proteins intended for use as ELISA standards are often supplied with stabilizers such as BSA to maintain protein integrity and minimize adsorption to plasticware. Carrier-free formulations may be less stable and more prone to loss during dilution steps.

• Calibration: Recombinant proteins are not inherently calibrated for ELISA quantification. Their mass must be value-assigned by measurement in the specific ELISA system, rather than relying solely on the mass stated on the vial label. This is because immunologically recognizable mass may differ between lots, and large dilution steps can introduce error.

• Validation: To use a recombinant protein as a standard, you must generate a standard curve in your ELISA using serial dilutions of the protein. The accuracy of quantification depends on the parallelism between the standard curve and the sample response. It is recommended to validate the standard curve with known concentrations and ensure consistent recovery and linearity.

• Assay Compatibility: The recombinant PDGF R-β must be recognized by the capture and detection antibodies used in your ELISA. If your assay is designed for native PDGF R-β, confirm that the recombinant form (including any tags or fusion partners) is immunologically equivalent in your system.

Best practices:

• Assign the concentration of your recombinant standard by running it in your ELISA and interpolating from a validated standard curve.
• Use the same buffer and matrix for standards and samples to minimize matrix effects.
• Validate the standard curve for linearity, sensitivity, and reproducibility in your assay.
• Document any lot-to-lot variability and re-validate if switching protein lots.

Summary Table: Recombinant Protein as ELISA Standard

In conclusion: You can use recombinant Mouse PDGF R-β as a standard for ELISA quantification, but you must calibrate and validate it within your specific assay system to ensure accurate and reproducible results.

Recombinant Mouse PDGF R-β has been validated for several key applications in published research, primarily as a tool for studying receptor-ligand interactions, signaling pathways, and as a standard or control in biochemical assays.

Validated Applications:

• Cell-based Functional Assays:
  Recombinant Mouse PDGF R-β is commonly used to study its ability to bind PDGF ligands and modulate downstream signaling. For example, its biological activity has been validated by its capacity to inhibit PDGF-BB-induced [^3H]-thymidine incorporation in NR6R-3T3 fibroblast cell lines, demonstrating its use in proliferation assays and receptor-ligand competition studies.

• ELISA (Enzyme-Linked Immunosorbent Assay):
  The recombinant protein is used as a standard or control in ELISA assays to quantify PDGF R-β levels in biological samples, and to validate antibody specificity for both natural and recombinant forms.

• Western Blot:
  Recombinant Mouse PDGF R-β is used as a positive control to validate antibody specificity and to assess protein expression in cell and tissue lysates.

• Blocking/Neutralization Studies:
  The recombinant receptor can be used to block or neutralize PDGF ligands in vitro, allowing researchers to dissect the functional consequences of PDGF signaling in various cell types.

• Receptor-Ligand Binding Studies:
  It is employed to characterize the binding affinities and specificities of PDGF isoforms (such as PDGF-AA, -BB, -CC, -DD) to the PDGF R-β, and to study receptor dimerization and activation mechanisms.

Additional Context:

• Signaling Pathway Analysis:
  Recombinant PDGF R-β is used to investigate downstream signaling events, such as activation of MAPK, PI3K, and PLCγ pathways, following ligand binding and receptor dimerization.

• Cellular and Molecular Biology:
  The protein is a valuable reagent for dissecting the roles of PDGF signaling in cell proliferation, migration, survival, and differentiation, particularly in fibroblasts, smooth muscle cells, and pericytes.

• Immunohistochemistry/Immunofluorescence:
  While the recombinant protein itself is not directly used for staining, antibodies validated against recombinant PDGF R-β are used in IHC and IF to detect endogenous receptor expression in tissues.

Summary Table of Validated Applications

If you need protocol details or specific examples from primary literature, please specify the application of interest.

Citations:
Western blot, IHC, and qPCR validation
Functional inhibition, receptor-ligand studies, signaling pathway analysis
ELISA, blocking/neutralization, and control applications
Immunofluorescence validation
Signaling and cellular function studies

To reconstitute and prepare Recombinant Mouse PDGF R-β protein for cell culture experiments, dissolve the lyophilized protein in sterile phosphate-buffered saline (PBS) at a concentration of 500 μg/mL. Centrifuge the vial briefly before opening to ensure all material is at the bottom.

Step-by-step protocol:

• Centrifuge the vial briefly before opening to collect the powder at the bottom.
• Reconstitute the lyophilized protein in sterile PBS to a final concentration of 500 μg/mL. If a different concentration is required, adjust the volume accordingly.
• If recommended by the manufacturer or for enhanced stability, you may add a carrier protein such as 0.1–1% bovine serum albumin (BSA) or fetal bovine serum (FBS) to the buffer.
• Aliquot the reconstituted protein into working volumes to avoid repeated freeze-thaw cycles.
• Store aliquots at –20°C to –70°C in a manual defrost freezer. Avoid frost-free freezers and repeated freeze-thaw cycles.
• For short-term use, storage at 2–8°C for up to one month is permissible under sterile conditions.
• Make further dilutions for cell culture in sterile, low-endotoxin medium or buffer containing carrier protein.

Additional notes:

• Always consult the specific product datasheet for any unique requirements, such as buffer composition or recommended reconstitution concentration.
• Confirm the absence of calcium and magnesium in the PBS if specified, as these ions can affect protein stability or activity.
• The optimal working concentration for cell culture should be determined empirically for your specific assay.
• Ensure all solutions and materials are sterile to prevent contamination.

Summary of best practices:

• Use sterile PBS for initial reconstitution.
• Add carrier protein if recommended or if protein stability is a concern.
• Aliquot and store at –20°C to –70°C.
• Avoid repeated freeze-thaw cycles.
• Prepare final working dilutions in cell culture medium immediately before use.

These steps will help maintain the integrity and biological activity of Recombinant Mouse PDGF R-β for cell culture experiments.