Recombinant Human Growth differentiation factor 11 (GDF-11)

Recombinant Human Growth Differentiation Factor 11 (GDF-11) is used in research applications primarily because of its demonstrated roles in tissue regeneration, reversal of age-related pathologies, and modulation of cellular processes relevant to aging, neurodegeneration, cardiovascular disease, and inflammation.

Key scientific reasons to use recombinant GDF-11 in research:

• Regeneration and Repair: GDF-11 stimulates regeneration in multiple tissues and organs, including the brain, heart, and skeletal muscle. It has been shown to promote neurogenesis, angiogenesis, and synaptic density, which are critical for recovery after injury such as stroke or neurodegenerative conditions.

• Reversal of Age-Related Pathologies: Supplementation with GDF-11 can reverse cardiac hypertrophy, bone loss, and pulmonary dysfunction in aged animal models, suggesting a rejuvenating effect on multiple organ systems.

• Neuroprotection and Functional Recovery: Recombinant GDF-11 reduces brain inflammation, atrophy, and gliosis, improves white matter integrity, and enhances locomotor activity after ischemic stroke or intracerebral hemorrhage. It also protects against mitochondrial dysfunction and oxidative stress in neuronal models.

• Immunomodulation: GDF-11 facilitates the conversion of pro-inflammatory M1 macrophages to anti-inflammatory M2 macrophages, thereby decreasing inflammatory responses. It also regulates macrophage metabolism and cholesterol removal, which may be relevant in studies of inflammation and metabolic disease.

• Muscle and Vascular Effects: GDF-11 increases skeletal muscle volume and strength, improves muscle tone, and enhances vascular regeneration, making it valuable for studies on muscle aging, sarcopenia, and vascular biology.

• Developmental Biology and Hematopoiesis: GDF-11 is involved in embryonic development and normal erythropoiesis, and its dysregulation is implicated in diseases such as anemia and myelodysplastic syndromes.

Experimental applications include:

• Investigating mechanisms of tissue regeneration and repair.
• Modeling age-related diseases and testing interventions for rejuvenation.
• Studying neuroprotection, neurogenesis, and recovery after brain injury.
• Exploring immunomodulatory and anti-inflammatory pathways.
• Assessing effects on muscle and vascular biology.

Considerations:

• There are some contradictory reports regarding the rejuvenating effects of GDF-11, so experimental design should include appropriate controls and consider dose-response relationships.
• Recombinant GDF-11 enables precise, reproducible studies by providing a defined, bioactive protein for in vitro and in vivo experiments.

In summary: Use of recombinant GDF-11 in research is justified by its multifaceted roles in regeneration, aging, neuroprotection, immunomodulation, and developmental biology, making it a valuable tool for investigating mechanisms and potential therapies for a wide range of conditions.

Yes, you can use recombinant Human Growth Differentiation Factor 11 (GDF-11) as a standard for quantification or calibration in ELISA assays, provided it is compatible with your assay system and matches the native protein's immunoreactivity.

Essential context and supporting details:

• ELISA Standard Preparation: Most commercial GDF-11 ELISA kits use recombinant human GDF-11 as the standard for generating calibration curves. The standard is typically reconstituted and serially diluted to create a range of known concentrations (e.g., 1000 pg/mL down to 15.6 pg/mL), which are then used to plot a standard curve for quantification.
• Assay Principle: The concentration of GDF-11 in unknown samples is determined by comparing their optical density (OD) readings to the standard curve generated using the recombinant protein.
• Specificity and Compatibility: It is important that the recombinant GDF-11 used as a standard is recognized by the antibodies in your ELISA kit and mimics the native protein's epitopes. Most kits specify that their standards and immunogens are recombinant proteins, and validation data typically show no significant cross-reactivity with analogues.
• Best Practices:
  • Prepare the standard according to the kit or protocol instructions, ensuring proper dilution and mixing to avoid foaming and ensure complete dissolution.
  • Always run the standard curve in parallel with your samples for each assay to account for potential variability.
  • Confirm the detection range and sensitivity of your assay to ensure your sample concentrations fall within the calibrated range.
  • If using a recombinant standard from a different source than your kit, verify its purity, sequence, and biological activity to ensure accurate quantification.

Additional relevant information:

• Validation: If you are developing your own ELISA or using a non-kit recombinant standard, perform a preliminary experiment to confirm that the recombinant GDF-11 produces a reliable standard curve and is detected equivalently to native GDF-11 in your samples.
• Documentation: Record the lot number, concentration, and preparation details of your recombinant standard for reproducibility and troubleshooting.

In summary, recombinant human GDF-11 is widely accepted as a standard for ELISA quantification, but ensure it is validated for your specific assay system and matches the immunoreactivity of the native protein.

Recombinant Human Growth Differentiation Factor 11 (GDF-11) has been validated in published research for several key applications, primarily in the fields of regenerative medicine, neuroscience, cardiovascular biology, oncology, and immunology.

Validated Applications:

• Neurogenesis and CNS Regeneration

  • GDF-11 enhances hippocampal neurogenesis, increases neural stem cell populations, improves vasculature, and boosts neuronal activity and plasticity in aged mice. It acts on brain endothelial cells to mediate these effects, rather than crossing the blood-brain barrier directly.
  • It promotes recovery after ischemic stroke by enhancing neovascularization, reducing inflammation, and improving sensorimotor function in rat models.

• Cardiac Hypertrophy and Fibrosis

  • Recombinant GDF-11 reduces pathological cardiac hypertrophy, decreases cardiac fibrosis, and improves cardiac pump function in pressure overload models.
  • It has also been shown to protect against myocardial damage in various cardiac injury models, including myocardial infarction and radiation-induced damage.

• Endothelial Cell Function and Vascular Biology

  • GDF-11 improves endothelial cell function, promotes enrichment of endothelial progenitor cells, and accelerates wound healing in diabetic models.
  • It alleviates endothelial dysfunction in atherosclerosis and diabetic retinopathy models.

• Cancer Biology

  • GDF-11 is involved in the differentiation of stem cells and is implicated in the progression of several cancers, including colon, breast, melanoma, oral squamous cell carcinoma, liver, pancreatic, esophageal cancer, and cholangiocarcinoma.

• Immunomodulation

  • GDF-11 facilitates the conversion of pro-inflammatory M1 macrophages to anti-inflammatory M2 macrophages, thereby reducing inflammatory responses and promoting metabolic regulation in macrophages.

• Developmental Biology

  • GDF-11 is essential for embryonic development, including neural, mesodermal, and olfactory tissue differentiation.

• Skeletal Muscle Regeneration

  • GDF-11 has been reported to increase skeletal muscle volume and enhance muscle strength, suggesting a role in muscle regeneration and anti-aging research.

Experimental Formats:

• Recombinant human GDF-11 has been used in both in vitro (cell culture) and in vivo (animal model) experiments, with validated protocols for administration and dosing.

Summary Table:

Note: Some studies report conflicting effects of GDF-11 in cardiac injury models, with certain research indicating exacerbation of myocardial injury under specific conditions. This highlights the importance of context, dosing, and experimental design in interpreting GDF-11’s biological effects.

In summary, recombinant human GDF-11 has been validated for use in neuroregeneration, cardiac protection, vascular biology, cancer research, immunomodulation, developmental biology, and muscle regeneration, with both in vitro and in vivo protocols established in the literature.

To reconstitute and prepare Recombinant Human Growth Differentiation Factor 11 (GDF-11) for cell culture experiments, follow these best-practice steps based on current protocols and technical datasheets:

1. Reconstitution Buffer and Concentration

• Preferred buffer: Use sterile 4 mM HCl for reconstitution, as this maintains protein solubility and stability.
• Alternative buffers: Some protocols allow reconstitution in sterile deionized water or 10 mM HCl. However, acidic conditions (HCl) are generally recommended to prevent aggregation.
• Concentration: Reconstitute to a final concentration of 0.1–1.0 mg/mL (100–1000 μg/mL), with 100–200 μg/mL being commonly used for stock solutions.

2. Reconstitution Procedure

• Briefly centrifuge the vial before opening to collect all lyophilized material at the bottom.
• Add the calculated volume of sterile 4 mM HCl (or other specified buffer) directly to the vial.
• Gently swirl or invert the vial to dissolve the protein. Do not vortex, as this may denature the protein.
• Allow the solution to sit at room temperature for 10–30 minutes to ensure complete dissolution.

3. Aliquoting and Storage

• Once fully dissolved, aliquot the solution into small volumes to avoid repeated freeze-thaw cycles, which can degrade the protein.
• Store aliquots at –20°C to –70°C for long-term storage. For short-term use (up to a few days), 2–8°C is acceptable.
• If desired, add a carrier protein (e.g., 0.1% BSA) or 5–50% glycerol to improve stability during storage, especially for dilute solutions.

4. Preparation for Cell Culture

• Before use, dilute the reconstituted stock to the desired working concentration using cell culture medium or other appropriate buffer.
• If the protein was reconstituted in acidic buffer (HCl), ensure the final dilution into cell culture medium is sufficient to neutralize the acid and avoid cytotoxicity.
• Filter sterilize the working solution if sterility is required and not already ensured by the reconstitution process (use a 0.2 μm filter).

5. Additional Notes

• Avoid repeated freeze-thaw cycles by using single-use aliquots.
• Do not vortex the protein solution at any stage.
• If the protein is tagged or fused (e.g., His-tag, SUMO-tag), confirm that the tag does not interfere with your downstream application.

Summary Table: GDF-11 Reconstitution Protocols

These steps will ensure maximum stability and bioactivity of recombinant GDF-11 for your cell culture experiments.