Recombinant Human PDGF-CC

Recombinant Human PDGF-CC is used in research applications because it is a potent growth factor involved in cell proliferation, migration, angiogenesis, tissue regeneration, and neuroprotection, making it valuable for studies in developmental biology, regenerative medicine, oncology, and vascular biology.

Key scientific applications and rationale include:

• Angiogenesis and Vascular Biology: PDGF-CC is a strong angiogenic factor, stimulating blood vessel growth and smooth muscle cell proliferation, which is critical for cardiovascular development and tissue repair studies.
• Tissue Regeneration and Wound Healing: PDGF family members, including PDGF-CC, promote recruitment and proliferation of cells essential for tissue repair, such as fibroblasts and osteoprogenitor cells, and stimulate extracellular matrix production. This makes it useful for modeling and enhancing wound healing, bone regeneration, and soft tissue repair.
• Neuroprotection: Recombinant PDGF-CC has demonstrated neuroprotective effects in retinal degeneration models, reducing apoptosis and stabilizing photoreceptor cells without inducing unwanted angiogenesis, which is relevant for neurodegenerative disease research.
• Cancer and Tumor Biology: PDGF-CC is expressed in various tumors and contributes to tumorigenesis and tumor-associated angiogenesis, making it a target for cancer biology and anti-angiogenic therapy studies.
• Developmental Biology: PDGF-CC is involved in embryonic development processes such as ductal morphogenesis and neuron development, supporting its use in developmental studies.
• Cell Signaling Studies: PDGF-CC activates specific PDGF receptors (notably PDGFR-α), allowing investigation of receptor-specific signaling pathways in different cell types.

Best practices for using recombinant PDGF-CC in research:

• Use in controlled bioassays to study cell proliferation, migration, and differentiation.
• Apply in in vitro and in vivo models of tissue injury, regeneration, or disease to assess functional outcomes.
• Combine with other growth factors or inhibitors to dissect signaling pathways and cellular responses.

In summary, recombinant human PDGF-CC is a versatile tool for dissecting mechanisms of cell growth, tissue repair, angiogenesis, and neuroprotection, and for modeling pathological processes such as fibrosis and cancer.

Yes, recombinant human PDGF-CC can be used as a standard for quantification or calibration in ELISA assays, provided it is compatible with your assay system. This is a common practice in quantitative ELISA protocols for growth factors and cytokines.

Key points and supporting details:

• ELISA kits for PDGF-CC typically use recombinant human PDGF-CC as the standard to generate the calibration curve against which unknown sample concentrations are interpolated. The standard is serially diluted to create a range of known concentrations, and the optical density (OD) readings from these standards are used to construct the standard curve.

• Assay specificity and compatibility: Most commercial human PDGF-CC ELISA kits are validated to detect both native and recombinant human PDGF-CC, and the standard curves generated with recombinant protein are shown to be parallel to those obtained with natural PDGF-CC. This indicates that recombinant PDGF-CC is suitable for accurate quantification in these assays.

• Protocol considerations:

  • Ensure the recombinant PDGF-CC is reconstituted and diluted in the same buffer as recommended by your ELISA protocol to avoid matrix effects or inaccuracies.
  • The concentration range of your standard curve should match the expected concentration range of your samples for optimal accuracy.
  • If using a custom or in-house ELISA, confirm that the antibodies used recognize the recombinant form equivalently to the native protein.

• Validation: If you are using a recombinant PDGF-CC standard from a source different from your ELISA kit, it is advisable to validate its performance by checking for parallelism between the standard curve generated with your recombinant protein and the kit’s standard, if available.

• Research use only: Most recombinant standards and ELISA kits are for research use only and not for diagnostic applications.

Summary Table: Recombinant PDGF-CC as ELISA Standard

In conclusion: Recombinant human PDGF-CC is widely used as a standard for quantification in ELISA assays, provided it is properly validated and matched to the assay system.

Recombinant Human PDGF-CC has been validated for several applications in published research, primarily in functional assays, bioassays, and studies of cell signaling, angiogenesis, neuroprotection, and barrier integrity.

Key validated applications include:

• Functional Assays and Bioassays:
  Recombinant PDGF-CC is widely used in functional assays to study its biological activity, such as stimulating cell proliferation, migration, and survival, particularly in smooth muscle cells, fibroblasts, and mesenchymal stem cells. It is also used in bioassays to assess its effects on various cell types and signaling pathways.

• Angiogenesis and Tumor Biology:
  PDGF-CC has been validated in angiogenesis assays (e.g., mouse cornea pocket assay, CAM assay) to study vessel growth and its role in tumor microenvironment modulation, including recruitment of cancer-associated fibroblasts and resistance to anti-VEGF therapies.

• Blood-Brain and Blood-Retinal Barrier Studies:
  Recombinant PDGF-CC is used to model and investigate blood-brain barrier (BBB) and blood-retinal barrier (BRB) disruption in vivo, as well as to test therapeutic interventions targeting these barriers in models of stroke, multiple sclerosis, traumatic brain injury, and neurodegeneration.

• Neuroprotection and CNS Disease Models:
  PDGF-CC has been validated in neuroprotection assays, including studies of neuronal survival after injury (e.g., optic nerve crush), and in models of HIV-associated neurocognitive disorders, where it modulates neurotrophic and apoptotic gene expression.

• Protein-Protein Interaction and Binding Assays:
  Used in binding assays and protein-protein interaction studies to characterize its interaction with PDGFRα and other signaling molecules.

• Cellular Signaling Studies:
  Recombinant PDGF-CC is employed to activate PDGFRα signaling in vitro, enabling the study of downstream pathways such as PI3K, TRPC, and GSK3β in various cell types.

• Cardiovascular Research:
  Validated for stimulating proliferation of coronary artery smooth muscle cells and studying cardiovascular development.

• Immunological and Expression Studies:
  Used as a standard or positive control in ELISA, immunohistochemistry, and Western blot for detecting PDGF-CC or its effects, though these are less frequently reported than functional assays.

Summary Table: Validated Applications for Recombinant Human PDGF-CC

These applications are supported by both primary research articles and product validation data from major suppliers, reflecting broad utility in cell biology, neuroscience, oncology, and vascular research.

To reconstitute and prepare Recombinant Human PDGF-CC protein for cell culture experiments, dissolve the lyophilized protein at 100 μg/mL in sterile 4 mM HCl, optionally containing at least 0.1% human or bovine serum albumin (HSA/BSA) as a carrier protein. This ensures protein stability and minimizes adsorption to surfaces.

Step-by-step protocol:

• Centrifuge the vial briefly before opening to collect all powder at the bottom.
• Add sterile 4 mM HCl to achieve a final concentration of 100 μg/mL. If recommended, include 0.1% HSA or BSA to further stabilize the protein, especially for long-term storage or repeated freeze-thaw cycles.
• Gently mix by pipetting up and down or swirling. Avoid vigorous vortexing to prevent protein denaturation.
• Allow the solution to sit for several minutes at room temperature to ensure complete dissolution.
• Aliquot the reconstituted protein into sterile tubes to avoid repeated freeze-thaw cycles.
• Storage after reconstitution:
  • Short-term: Store at 2–8 °C for up to 1 month under sterile conditions.
  • Long-term: Store at –20 °C to –70 °C for up to 3 months under sterile conditions.
  • Always avoid repeated freeze-thaw cycles.

Dilution for cell culture:

• Dilute the stock solution into your cell culture medium to the desired working concentration immediately before use. Use sterile, low-endotoxin buffers and media for all dilutions.
• If using serum-free media, ensure the presence of a carrier protein (e.g., BSA) to maintain protein stability.

Additional notes:

• If your protocol or supplier recommends reconstitution in sterile water instead of HCl, this is also acceptable, but stability may be reduced unless a carrier protein is present.
• Always consult the specific product datasheet for any unique instructions regarding your batch or formulation.

Summary of best practices:

• Reconstitute at 100 μg/mL in sterile 4 mM HCl ± 0.1% HSA/BSA.
• Aliquot and store at recommended temperatures.
• Avoid freeze-thaw cycles.
• Dilute freshly into cell culture medium before use.

These steps will help ensure the bioactivity and stability of PDGF-CC for your cell culture experiments.