Recombinant Mouse Persephin

Recombinant Mouse Persephin is used in research applications primarily because it is a neurotrophic factor that selectively promotes the survival and growth of central dopaminergic and motor neurons, making it valuable for studies in neurobiology, neurodegeneration, and kidney development.

Key reasons to use recombinant mouse Persephin in research include:

• Neurotrophic Activity: Persephin is structurally related to GDNF and neurturin, but is distinct in its selectivity. It supports the survival of ventral midbrain dopaminergic neurons and motor neurons, both in vitro and in vivo, but does not support peripheral neurons. This specificity is useful for dissecting pathways involved in central nervous system development, neuroprotection, and neurodegenerative disease models such as Parkinson’s disease.

• Kidney Development Studies: Persephin, like GDNF, promotes ureteric bud branching, making it relevant for developmental biology research focused on kidney morphogenesis.

• Defined, Reproducible Bioactivity: Recombinant Persephin is produced in controlled systems (e.g., E. coli), ensuring high purity and batch-to-batch consistency, which is critical for reproducible experimental results. Purity is typically >90% and endotoxin levels are kept low, suitable for sensitive cell-based assays.

• Versatile Applications: Recombinant Persephin can be used in:

  • Neuronal cell culture to assess survival, differentiation, or neuroprotection.
  • Kidney organoid or branching morphogenesis assays.
  • ELISA standards or as a control in biochemical assays.
  • Mechanistic studies of RET receptor signaling, as Persephin is a ligand for the RET tyrosine kinase receptor, though it may utilize distinct co-receptors compared to GDNF and neurturin.

• Research Use Only: Recombinant Persephin is intended for research purposes, not for diagnostic or therapeutic use, ensuring compliance with laboratory standards.

In summary, recombinant mouse Persephin is a powerful tool for studying central neuron survival, neurodegenerative disease mechanisms, and kidney development due to its selective bioactivity, high purity, and reproducibility in experimental systems.

You can use recombinant Mouse Persephin as a standard for quantification or calibration in ELISA assays, provided it is of high purity and its concentration is accurately determined. This is a common practice in ELISA development and commercial kits, where recombinant proteins are used to generate standard curves for quantification.

Key considerations:

• Purity and Characterization: The recombinant Persephin should be highly purified (typically >95%) and well-characterized, ideally with confirmation of its identity and activity.
• Concentration Accuracy: The protein concentration must be precisely measured, as errors here will directly affect the accuracy of your standard curve and quantification.
• Formulation: Recombinant standards are often supplied lyophilized and may contain stabilizers such as BSA or trehalose. If your recombinant Persephin contains carriers or additives, ensure they do not interfere with your assay.
• Matrix Effects: If your samples are in a complex matrix (e.g., serum, plasma), ensure that the recombinant standard is diluted in a similar matrix or in a suitable diluent to minimize matrix effects and improve accuracy.
• Validation: It is important to validate that your recombinant Persephin behaves similarly to endogenous Persephin in your assay system. Some ELISA kits specify that their standards are recombinant proteins, and validation data (such as recovery and linearity) are provided.

Caveats:

• Some ELISA kits or protocols may not recommend using recombinant proteins as standards if the assay is not validated for them, or if the recombinant protein differs in post-translational modifications or folding compared to the native protein. Always check the compatibility with your specific ELISA system.
• If you are using a commercial ELISA kit, it is best to use the standard provided with the kit, as it is validated for that specific assay.

Summary Table:

In summary, recombinant Mouse Persephin is suitable as a standard for ELISA quantification if it is pure, accurately quantified, and validated for your assay conditions. Always confirm compatibility with your specific ELISA protocol or kit.

Recombinant Mouse Persephin has been validated primarily for bioactivity assays, with published research supporting its use in several specific applications related to neurobiology and cell biology.

Key validated applications include:

• Bioactivity/Functional Assays: Persephin has been shown to promote the survival of ventral midbrain dopaminergic neurons and motor neurons in both in vitro and in vivo models, particularly after injury or neurotoxic insult. This is the principal application validated in both product datasheets and primary literature.
• Cell Proliferation Assays: Some commercial sources specifically list cell proliferation assays as a validated application, indicating its use in assessing the effects of Persephin on neuronal or other cell types in culture.
• ELISA (Enzyme-Linked Immunosorbent Assay): Recombinant Persephin is used as a standard or control in ELISA protocols to quantify protein levels or validate antibody specificity.
• Western Blot: It is also validated for use as a positive control in Western blotting to detect Persephin or related signaling components.
• Kidney Development Studies: Persephin has been used in studies of ureteric bud branching, reflecting its role in kidney morphogenesis.
• Neuroprotection/Neurodegeneration Models: Published research demonstrates its use in models of neurodegenerative diseases, such as Parkinson’s disease, to assess neuroprotective effects.

Summary Table of Validated Applications

Additional Notes:

• Most applications are research-focused and not for diagnostic or therapeutic use.
• The protein is typically supplied as a lyophilized, E. coli-derived product, validated for high purity and low endotoxin content.
• Published research highlights its specificity for central neurons (dopaminergic and motor), with little to no effect on peripheral neurons.

If you require protocols or more detailed application notes for a specific assay, please specify the intended use.

To reconstitute and prepare Recombinant Mouse Persephin protein for cell culture experiments, follow these steps to ensure protein stability and biological activity:

1. Equilibrate and Centrifuge

   • Allow the lyophilized vial and your reconstitution buffer (commonly sterile 4 mM HCl or sterile water, as specified in the product datasheet) to reach room temperature.
   • Briefly centrifuge the vial to collect all powder at the bottom.

2. Reconstitution

   • Add sterile 4 mM HCl to the vial to achieve a final concentration of 0.1–1.0 mg/mL (e.g., for 100 µg, add 1 mL for 0.1 mg/mL or 100 µL for 1 mg/mL).
   • Gently swirl or invert the vial to dissolve the protein. Do not vortex or shake vigorously, as this can denature the protein.
   • Let the solution stand at room temperature for 15–30 minutes with gentle agitation to ensure complete dissolution.

3. Dilution for Cell Culture

   • Once fully dissolved, dilute the reconstituted Persephin to your desired working concentration using cell culture medium or buffer containing a carrier protein such as 0.1% BSA, 5% HSA, or 10% FBS to prevent adsorption and loss of activity.
   • If your experiment requires serum-free conditions, avoid animal-derived carriers and consider using trehalose as a stabilizer.

4. Aliquot and Storage

   • Aliquot the solution to avoid repeated freeze-thaw cycles, which can degrade the protein.
   • Store aliquots at –20°C to –80°C for long-term storage. For short-term use (up to one week), store at 2–8°C.
   • For long-term storage, adding 5–50% glycerol or a carrier protein can further stabilize the protein.

5. Usage Notes

   • Use sterile technique throughout to prevent contamination.
   • Always consult the specific product datasheet or Certificate of Analysis for any unique requirements for your Persephin preparation, as formulations may vary between suppliers.

Summary Table: Key Steps for Recombinant Mouse Persephin Reconstitution

These steps will help maintain the biological activity and stability of Recombinant Mouse Persephin for your cell culture experiments.