Recombinant Human PDGF-AA

Recombinant Human PDGF-AA is widely used in research applications due to its potent biological activities, particularly its ability to stimulate cell proliferation, migration, and differentiation in various cell types.

Key scientific reasons to use Recombinant Human PDGF-AA:

• Mitogenic Activity: PDGF-AA is a strong mitogen for fibroblasts, smooth muscle cells, and connective tissue cells, making it valuable for studies involving cell growth, wound healing, and tissue regeneration.
• Chemotactic Effects: It attracts fibroblasts, smooth muscle cells, neutrophils, and mononuclear cells, supporting research in cell migration, inflammation, and tissue remodeling.
• Stem Cell Differentiation: PDGF-AA is commonly used to differentiate human pluripotent stem cell-derived neural progenitor cells into oligodendrocyte precursor cells, which is critical for neural development and remyelination studies.
• Regenerative Medicine: PDGF-AA signaling is involved in the development of cranial and cardiac neural crest, lung, intestine, skeleton, and vasculogenesis, making it relevant for studies in tissue engineering and regenerative therapies.
• Neurobiology: PDGF-AA plays a role in neuron survival, regeneration, and glial cell proliferation and differentiation, supporting research in neurodevelopment and neurodegenerative diseases.
• Immunomodulation: It can transiently induce T cell IL-2 secretion and modulate cytokine production, which is useful for immunology research.
• Bone and Vascular Biology: PDGF-AA is implicated in bone regeneration, vascular remodeling, and mesenchymal stromal cell heterogeneity, making it suitable for studies in bone healing and vascular biology.

Typical research applications include:

• Cell culture assays to assess proliferation, migration, and differentiation.
• Stem cell maintenance and lineage specification, especially in neural and mesenchymal contexts.
• Functional assays in tissue engineering, wound healing, and regenerative medicine.
• Studies of vascular biology, fibrosis, and pathological mesenchymal responses.

Summary:
Use Recombinant Human PDGF-AA in your research if you require a defined, reproducible growth factor to stimulate proliferation, migration, or differentiation of mesenchymal, neural, or vascular cells, or to model regenerative, developmental, or pathological processes in vitro or in vivo.

You can use recombinant human PDGF-AA as a standard for quantification or calibration in your ELISA assays, provided that the recombinant protein is of high purity, its concentration is accurately known, and it is compatible with your assay system.

Key considerations and supporting details:

• ELISA standards are typically recombinant proteins: Most commercial human PDGF-AA ELISA kits use recombinant human PDGF-AA as the standard to generate the calibration curve for quantification. The standard curve is constructed by measuring known concentrations of recombinant PDGF-AA, and sample concentrations are interpolated from this curve.

• Parallelism and comparability: Studies and kit documentation indicate that recombinant human PDGF-AA produces standard curves that are parallel to those generated with natural PDGF-AA, supporting its use for accurate quantification. This parallelism is essential to ensure that the recombinant standard behaves similarly to the native protein in your samples.

• Purity and quantification: The recombinant PDGF-AA used as a standard should be highly purified and its concentration precisely determined, typically by absorbance at 280 nm or amino acid analysis. Impurities or inaccurate quantification of the standard will directly affect the accuracy of your ELISA results.

• Matrix effects: When using a recombinant standard, it is important to prepare the standard curve in a matrix similar to your samples (e.g., serum, plasma, or the same buffer) to minimize matrix effects that could alter antigen-antibody interactions. Some protocols recommend spiking the recombinant standard into a sample matrix to better match the sample environment.

• Documentation and validation: Always consult your specific ELISA kit protocol to confirm compatibility. Some kits are validated only with their supplied standards, and using an external recombinant standard may require additional validation to ensure accuracy and reproducibility.

Best practices:

• Use recombinant PDGF-AA of known concentration and high purity.
• Prepare the standard curve in a matrix similar to your samples.
• Validate parallelism between the standard curve and sample dilution curves.
• Follow the ELISA kit manufacturer’s recommendations regarding standards and controls.

In summary, recombinant human PDGF-AA is widely accepted and used as a standard for ELISA quantification, provided you ensure purity, accurate quantification, and matrix compatibility.

Recombinant Human PDGF-AA has been validated in published research for a range of applications, primarily involving its roles as a mitogen, chemoattractant, and differentiation factor in various cell types and biological systems.

Key validated applications include:

• Cell Proliferation and Bioassays:
  PDGF-AA is widely used to stimulate proliferation in fibroblasts (e.g., NR6R-3T3 mouse fibroblast cell line), smooth muscle cells, and other mesenchymal cells in dose-dependent bioassays. It is also used to assess mitogenic activity in various cell-based functional assays.

• Cell Migration and Chemotaxis:
  It has been validated for studying chemotactic responses in fibroblasts, smooth muscle cells, neutrophils, and mononuclear cells.

• Stem Cell Maintenance and Differentiation:
  PDGF-AA is used to maintain and differentiate stem and progenitor cells, notably:

  • Differentiation of human pluripotent stem cell (hPSC)-derived neural progenitor cells into oligodendrocyte precursor cells.
  • Maintenance and differentiation of mesenchymal stem cells and immune cells.

• Neural and Glial Cell Studies:
  It is applied in research on neuron survival, regeneration, and glial cell proliferation/differentiation, reflecting its suggested roles in the central nervous system.

• Tissue Regeneration and Repair:
  Recombinant human PDGF-AA has been validated in models of bone regeneration, periodontal defect repair, and soft tissue healing, leveraging its ability to recruit osteoprogenitor cells and promote angiogenesis. It is FDA-approved for periodontal defect repair and is under investigation for broader skeletal and soft tissue applications.

• Immunomodulation:
  PDGF-AA has been used to study immune cell modulation, including effects on neutrophil granule release, phagocytosis, inhibition of NK cell activity, and transient modulation of T cell cytokine secretion.

• Vascular Biology:
  It is validated for studies on vascular smooth muscle cell growth, angiogenesis, and vascular remodeling.

• Functional and Biochemical Assays:
  PDGF-AA is used in ELISA, SDS-PAGE, and other biochemical assays as a protein standard or functional reagent.

Representative published research applications:

• Differentiation of iPS cells into oligodendrocyte precursors.
• Mesenchymal stromal cell heterogeneity and bone regeneration studies.
• Neurodevelopmental and neurodegenerative disease models.
• Vascular remodeling and neointima formation in chronic kidney disease models.
• CNS remyelination and neuroprotection studies.

Summary Table of Validated Applications

These applications are supported by both product validation data and peer-reviewed publications, reflecting the broad utility of recombinant human PDGF-AA in cell biology, regenerative medicine, and immunology research.

To reconstitute and prepare Recombinant Human PDGF-AA protein for cell culture experiments, dissolve the lyophilized protein in sterile water or a specified acidic buffer to a concentration of 0.1–1.0 mg/mL, then dilute further in cell culture medium containing a carrier protein such as 0.1% BSA or HSA to prevent adsorption and loss of activity.

Step-by-step protocol:

1. Centrifuge the vial briefly before opening to ensure all lyophilized material is at the bottom.
2. Reconstitution:
   • Add sterile, deionized water directly to the vial to achieve a concentration between 0.1–1.0 mg/mL (commonly 0.1 mg/mL is used).
   • For some formulations, reconstitution in 4 mM HCl at 100 μg/mL is recommended; always check the specific product datasheet for buffer requirements.
   • Do not vortex; gently pipette to mix until fully dissolved.
3. Aliquot and Storage:
   • Store the reconstituted stock at 2–8°C for up to 1 week.
   • For longer-term storage, aliquot and freeze at –20°C to –80°C. Avoid repeated freeze-thaw cycles.
   • For extended storage, dilute the stock in buffer containing 0.1% BSA or HSA to stabilize the protein.
4. Preparation for cell culture:
   • Prepare working dilutions in cell culture medium or PBS containing 0.1% BSA or HSA to minimize adsorption to plasticware and maintain bioactivity.
   • Typical working concentrations for cell stimulation range from 10–200 ng/mL, depending on cell type and experimental design.
5. Handling precautions:
   • Avoid vigorous mixing or vortexing, which can denature the protein.
   • Use sterile technique throughout to prevent contamination.

Summary Table:

Key notes:

• Always consult the specific product datasheet for buffer requirements and recommended concentrations.
• Carrier proteins (BSA/HSA) are essential for stability and preventing loss due to adsorption.
• Avoid repeated freeze-thaw cycles to maintain protein integrity.

This protocol ensures optimal solubility, stability, and biological activity of recombinant PDGF-AA for cell culture applications.