Recombinant Mouse PDGF-AA

Recombinant Mouse PDGF-AA is a valuable tool for research applications due to its well-characterized biological activity and broad utility in studying cellular processes. Here are several key reasons why you should consider using Recombinant Mouse PDGF-AA in your research:

1. Mitogenic Activity

Recombinant Mouse PDGF-AA is a potent mitogen for a wide range of cell types, including fibroblasts, smooth muscle cells, connective tissue cells, bone and cartilage cells, and some blood cells. This makes it ideal for studies involving cell proliferation and growth.

2. Cell Migration and Wound Healing

PDGF-AA plays a crucial role in directing cell migration during embryonic development and postnatal processes such as wound healing. Using recombinant PDGF-AA can help investigate mechanisms of cell migration and tissue repair.

3. Epithelial-Mesenchymal Interactions

PDGF-AA and its receptor are important in epithelial-mesenchymal interactions, which are critical for early lung branching morphogenesis. This makes it useful for developmental biology studies.

4. Regulation of VEGF Expression

PDGF-AA is an essential autocrine regulator for vascular endothelial growth factor (VEGF) expression, influencing angiogenesis and vascular development. This is particularly relevant for cancer research and studies on vascular disorders.

5. Neuronal Development and Survival

PDGF-AA is involved in embryonic neuron development and is suggested to play a role in neuron survival and regeneration in the central nervous system. This makes it suitable for neuroscience research.

6. High Purity and Consistency

Recombinant Mouse PDGF-AA is typically produced with high purity (>97% by SDS-PAGE and HPLC) and is available in a lyophilized form, ensuring consistency and reliability in experimental results.

7. Well-Characterized Biological Activity

The biological activity of recombinant PDGF-AA is well-established, with dose-dependent stimulation of cell proliferation (e.g., Balb/c 3T3 cells) and a known ED50 (8-10 ng/ml), allowing for precise dosing in experiments.

8. Versatility in Research Applications

PDGF-AA is involved in various biological processes, including hyperplasia, chemotaxis, and respiratory tubule epithelial cell development, making it a versatile reagent for multiple research areas.

9. Support for Gene Therapy Studies

Recombinant PDGF-AA can be used to study the effects of gene delivery and to modulate the activity of cells such as cementoblasts, providing insights into tissue regeneration and periodontal therapy.

10. Research Use Only

Recombinant Mouse PDGF-AA is intended for research use only, ensuring that it meets the regulatory requirements for laboratory studies and is not intended for human use.

In summary, Recombinant Mouse PDGF-AA is a powerful and versatile reagent that can enhance your research in cell biology, developmental biology, neuroscience, and tissue engineering. Its well-characterized activity and high purity make it a reliable choice for a wide range of experimental applications.

You can use recombinant Mouse PDGF-AA as a standard for quantification or calibration in ELISA assays, provided it is compatible with your assay's antibodies and detection system. This is a common practice in quantitative ELISA, where recombinant proteins serve as standards to generate a calibration curve against which sample concentrations are determined.

Key considerations and supporting details:

• Standard Curve Generation: ELISA kits for Mouse PDGF-AA typically use recombinant Mouse PDGF-AA to generate the standard curve. The concentration of PDGF-AA in unknown samples is calculated by comparing their optical density (OD) values to this standard curve.
• Assay Compatibility: Ensure that the recombinant Mouse PDGF-AA you use as a standard is recognized by the capture and detection antibodies in your specific ELISA kit. Most commercial kits are validated with recombinant standards, but cross-reactivity or epitope differences can affect quantification if the recombinant protein differs significantly from the native form.
• Validation: Some ELISA kits explicitly state that results obtained using natural PDGF-AA are parallel to those obtained with recombinant standards, supporting the use of recombinant proteins for calibration. However, it is good practice to verify parallelism between your recombinant standard and native samples, especially if you are using a custom or non-validated assay.
• Matrix Effects: Recovery and linearity studies in various sample matrices (e.g., serum, plasma, cell culture supernatant) are important to confirm that the recombinant standard behaves similarly to endogenous PDGF-AA in your assay system.
• Documentation: Follow the ELISA kit protocol for standard preparation, dilution, and curve fitting to ensure accurate quantification.

Summary Table: Use of Recombinant Mouse PDGF-AA as ELISA Standard

In conclusion, recombinant Mouse PDGF-AA is suitable as a standard for ELISA quantification if it is compatible with your assay system and validated for your application. Always consult your specific ELISA kit documentation for any restrictions or recommendations regarding standard selection.

Recombinant Mouse PDGF-AA has been validated in published research for several key applications, primarily as a biologically active growth factor in cell-based assays and as a control or standard in analytical techniques.

Validated Applications in Published Research:

• Cell Proliferation Assays: PDGF-AA is widely used to stimulate proliferation of mouse fibroblast cell lines (e.g., 3T3, NIH3T3) in vitro, serving as a positive control for mitogenic activity.
• Bioactivity Assays: Its ability to induce cellular responses such as proliferation, migration, and chemotaxis in mesenchymal-origin cells (fibroblasts, smooth muscle cells, glial cells) is routinely validated in bioassays.
• Western Blot Control: PDGF-AA is used as a protein standard or control in Western blot experiments to confirm antibody specificity and protein detection.
• ELISA Standard: It serves as a quantitative standard in ELISA assays for measuring PDGF-AA levels in biological samples.
• Stem/Immune Cell Differentiation: PDGF-AA has been used to differentiate stem cells, including human iPS cells into oligodendrocyte precursors, and to support maintenance or differentiation of various stem and immune cell types.
• Fibrosis and Muscle Regeneration Studies: In disease models such as Duchenne muscular dystrophy (DMD), PDGF-AA is used to stimulate fibro-adipogenic progenitor cells (FAPs) to study pathways involved in muscle fibrosis and regeneration, validated by proliferation assays, proteomics, Western blot, immunofluorescence, and flow cytometry.
• Developmental Biology: PDGF-AA is essential for normal embryonic development, including lung alveolar formation, gastrointestinal tract development, Leydig cell development, and myelination in the CNS; these roles are validated in mouse models and cell culture systems.
• Chemotaxis and Migration Assays: PDGF-AA is used to study cell migration and chemotaxis, particularly in mesenchymal and glial cells.
• Wound Healing Models: Its role in promoting cell proliferation and migration is leveraged in in vitro and in vivo wound healing studies.

Summary Table of Validated Applications

These applications are supported by published studies and product validation data, confirming the utility of recombinant mouse PDGF-AA in diverse experimental contexts.

To reconstitute and prepare Recombinant Mouse PDGF-AA protein for cell culture experiments, dissolve the lyophilized protein in sterile water at a concentration of 0.1 mg/mL (100 μg/mL) or higher. This stock solution can then be further diluted in cell culture medium or other aqueous buffers as needed for your experiment.

Step-by-step protocol:

• Centrifuge the vial briefly before opening to ensure all powder is at the bottom.
• Add sterile water (preferably 18 MΩ-cm grade) to achieve a concentration between 0.1–0.5 mg/mL (e.g., add 1 mL water to 0.1–0.5 mg protein).
• Gently pipette up and down and wash the sides of the vial to fully dissolve the protein. Do not vortex.
• If required, allow the solution to sit at room temperature for a few minutes to ensure complete dissolution.
• Aliquot the reconstituted protein to avoid repeated freeze-thaw cycles.
• For long-term storage, add a carrier protein such as 0.1% HSA or BSA to prevent adsorption and degradation, then store aliquots at –20°C or below.
• For short-term use, store at 4°C for up to 2–7 days.

Additional notes:

• Avoid reconstituting at concentrations lower than 100 μg/mL to maintain protein stability.
• Always use sterile technique to prevent contamination.
• After reconstitution, the protein can be diluted into cell culture medium or buffer for experimental use.
• PDGF-AA is a disulfide-linked homodimer and is typically expressed in E. coli as a non-glycosylated protein.
• Biological activity should be confirmed by dose-dependent stimulation of cell proliferation (e.g., Balb/c 3T3 cells), with typical ED50 values around 8–10 ng/mL.

Summary Table:

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