Recombinant Rat GDNF Rα1

Recombinant Rat GDNF Rα1 is used in research applications to study and manipulate the GDNF signaling pathway, which is critical for neuronal survival, differentiation, and regeneration, particularly in models of neurodegeneration and kidney development.

Key scientific reasons to use Recombinant Rat GDNF Rα1:

• Essential receptor component: GDNF Rα1 (GFRα1) is the primary ligand-binding receptor for GDNF, forming a complex with the RET tyrosine kinase to mediate GDNF’s biological effects. Without GFRα1, GDNF cannot efficiently signal, making recombinant GDNF Rα1 indispensable for in vitro and in vivo studies of GDNF function.
• Modeling neuroprotection: GDNF, via GFRα1, protects and restores dopaminergic neurons in the substantia nigra, which is highly relevant for Parkinson’s disease research and other neurodegenerative models. Recombinant GDNF Rα1 enables controlled activation of this pathway, allowing researchers to dissect mechanisms of neuronal survival and regeneration.
• Developmental biology: GFRα1 is crucial for kidney morphogenesis and the development of the enteric nervous system. Recombinant GDNF Rα1 is used to study these processes, as knockout models lacking GFRα1 show deficits similar to those lacking GDNF or RET, confirming its essential role.
• Ligand specificity and receptor preference: GDNF preferentially signals through GFRα1, and recombinant forms allow for precise manipulation of receptor-ligand interactions in cell-based assays, helping to clarify the specificity and cross-talk within the GDNF family.
• Functional assays: Recombinant GDNF Rα1 is used in binding studies, receptor activation assays, and to reconstitute signaling in cells or tissues lacking endogenous GFRα1, enabling mechanistic studies of GDNF-dependent processes.

Typical applications include:

• Neuroprotection and regeneration assays in rodent models of Parkinson’s disease and other neurodegenerative conditions.
• Developmental studies of the kidney and nervous system using knockout or rescue experiments.
• Cell signaling and receptor binding assays to dissect GDNF/GFRα1/RET interactions.
• In vitro differentiation and survival studies of neuronal cultures.

Best practices:

• Use recombinant GDNF Rα1 in combination with GDNF and RET to fully reconstitute the signaling pathway in cell-based or animal models.
• Validate the biological activity of recombinant protein in your specific assay system, as receptor-ligand interactions can be context-dependent.

In summary, Recombinant Rat GDNF Rα1 is a critical tool for dissecting GDNF-mediated signaling in neurobiology and developmental biology, enabling precise control and mechanistic understanding of this essential pathway.

Yes, recombinant rat GDNF (Glial cell line-derived neurotrophic factor) is commonly used as a standard for quantification or calibration in ELISA assays designed to measure rat GDNF. This practice is well-established in commercial ELISA kits, which routinely employ recombinant rat GDNF as the reference standard for generating calibration curves.

Key technical considerations:

• Standard Curve Generation: Recombinant rat GDNF is serially diluted to create a standard curve, which allows for the quantification of GDNF concentrations in unknown samples by interpolation.
• Assay Compatibility: Ensure that the recombinant protein matches the form of GDNF recognized by the antibodies in your ELISA (e.g., correct isoform, post-translational modifications if relevant). Most kits specify that their antibodies detect both natural and recombinant rat GDNF.
• Purity and Activity: The recombinant standard should be well-characterized, with known concentration and biological activity, to ensure accurate quantification.
• Storage and Handling: Follow recommended storage conditions (typically -20°C for lyophilized protein) and prepare fresh dilutions for each assay to maintain consistency and reliability.

Limitations and best practices:

• Use only recombinant rat GDNF that has been validated for ELISA applications, as differences in expression systems or purification methods can affect assay performance.
• Always prepare a fresh standard curve for each experiment to account for potential variability in protein stability or handling.
• Confirm that your ELISA kit or custom assay is optimized for recombinant rat GDNF as the standard, as some kits may have specific requirements or recommendations.

Summary Table: Use of Recombinant Rat GDNF as ELISA Standard

In conclusion, recombinant rat GDNF is suitable and widely accepted as a standard for ELISA quantification and calibration, provided it is properly validated and handled according to best laboratory practices.

Recombinant Rat GDNF Rα1 has been validated in published research primarily for bioassays, cell culture studies, and in vivo applications focused on neurobiology, stem cell biology, and neuroprotection.

Key validated applications include:

• Bioassays:
  Used to assess the biological activity of GDNF signaling, including its effects on neuronal survival, differentiation, and proliferation in various cell types (e.g., neuronal, glial, and stem cells).
  Examples:

  • Promotion of dopaminergic neuron survival and differentiation.
  • Induction of c-Fos transcription and proliferation in spermatogonial stem cells.
  • Regulation of neurotrophin receptor signaling and intracellular trafficking in neuronal cells.

• Cell Culture:
  Applied to cultured cells to study GDNF-mediated signaling pathways, neuroprotection, and cellular responses such as migration, survival, and gene expression changes.

• In Vivo Studies:
  Administered to live animals (rats and mice) to investigate neuroprotective effects, behavioral outcomes, and therapeutic potential in models of neurodegenerative diseases (e.g., Parkinson’s disease), nerve injury, and developmental studies.
  Examples:

  • In vivo administration to rats to study cold hyperalgesia after nerve injury.
  • Evaluation of GDNF’s role in neuromuscular junction development and maintenance.

• ELISA (Enzyme-Linked Immunosorbent Assay):
  Used as a standard or for assay development to quantify GDNF levels in biological samples.

Representative research areas:

• Neuroprotection: GDNF Rα1 has been used to demonstrate protection of dopaminergic neurons and prevention of motor neuron apoptosis.
• Stem Cell Biology: Studies on spermatogonial stem cell proliferation and migration in response to GDNF signaling.
• Neurodevelopment: Investigation of GDNF’s role in neuromuscular junction formation and maintenance.
• Behavioral Neuroscience: Assessment of learning, memory, and behavioral impairments following manipulation of GDNF levels in neonatal and adult rats.

Summary Table of Validated Applications

Note:
Most published validations focus on bioactivity assays (cell-based and in vivo), with additional use in ELISA and cell culture protocols. The protein is not validated for diagnostic or therapeutic use in humans.

If you require protocols or more specific experimental details for a particular application, please specify the context or research focus.

To reconstitute and prepare Recombinant Rat GDNF Rα1 protein for cell culture experiments, follow these steps for optimal protein stability and biological activity:

1. Reconstitution

• Concentration: Reconstitute the lyophilized protein at 100 μg/mL.
• Buffer: Use sterile PBS (phosphate-buffered saline), ideally containing at least 0.1% human or bovine serum albumin (BSA) as a carrier protein to prevent adsorption and loss of activity.
• Procedure:
  • Briefly centrifuge the vial to collect the lyophilized powder at the bottom before opening.
  • Add the appropriate volume of sterile PBS (with BSA) directly to the vial.
  • Gently pipette up and down or swirl to dissolve the protein completely. Avoid vigorous vortexing or foaming, which can denature the protein.

2. Aliquoting and Storage

• Aliquot the reconstituted protein into small volumes to avoid repeated freeze-thaw cycles, which can degrade the protein.
• Short-term storage: Store at 2–8 °C for up to 1 month under sterile conditions.
• Long-term storage: Store at –20 °C to –70 °C for up to 3 months. Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
• Do not store at –80 °C unless specified by the manufacturer, as some formulations may be sensitive to ultra-low temperatures.

3. Preparation for Cell Culture

• Working concentration: For most cell-based assays, the effective dose (ED₅₀) for GDNF is typically 1–3 ng/mL. However, optimal concentrations may vary depending on cell type and experimental design.
• Dilution: Dilute the reconstituted stock solution into your cell culture medium immediately before use. Use sterile technique throughout.
• Carrier protein: If not already present in your stock, ensure the final dilution contains a small amount of carrier protein (e.g., 0.1% BSA) to maintain stability.

4. Additional Notes

• Avoid repeated freeze-thaw cycles by aliquoting.
• Check for visible particulates after reconstitution; if present, clarify by brief centrifugation or filtration through a 0.22 μm sterile filter.
• Do not vortex the protein solution, as this can cause denaturation.

Summary Table: Key Steps for Recombinant Rat GDNF Rα1 Preparation

These guidelines ensure protein stability and activity for reliable cell culture experiments. Always consult the specific product datasheet for any additional instructions or formulation-specific recommendations.