Glial cell line-derived neurotrophic factor family receptor alpha-2, also known as GFRA2, is the cognate co-receptor for the neurotrophic factor neurturin that is selectively expressed by isolectin B(4) (IB 4)-binding nociceptive sensory neurons.1 GFRA2 is required for the development of the myenteric nerve plexus and is an excellent candidate gene for Hirschsprung disease (HSCR).2 GFRalpha2 signaling is also needed for development of the parasympathetic islet innervation that is critical for vagally induced hormone secretion.3
The predicted molecular weight of Recombinant Mouse GFRα-2 is Mr 74 kDa. However, the actual molecular weight as observed by migration on SDS-PAGE is Mr 90 kDa.
Predicted Molecular Mass
74
Formulation
This recombinant protein was 0.2 µm filtered and lyophilized from modified Dulbecco’s phosphate buffered saline (1X PBS) pH 7.2 – 7.3 with no calcium, magnesium, or preservatives.
Storage and Stability
This lyophilized protein is stable for six to twelve months when stored desiccated at -20°C to -70°C. After aseptic reconstitution, this protein may be stored at 2°C to 8°C for one month or at -20°C to -70°C in a manual defrost freezer. Avoid Repeated Freeze Thaw Cycles. See Product Insert for exact lot specific storage instructions.
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Recombinant Mouse GFRα-2 is used in research applications to study neurotrophic signaling, neuronal development, and disease mechanisms involving the GDNF (glial cell line-derived neurotrophic factor) family, particularly in the context of sensory neuron biology, enteric nervous system development, and neurodegenerative disease models.
Key reasons to use recombinant Mouse GFRα-2 in research:
Co-receptor Functionality: GFRα-2 is the primary co-receptor for the neurotrophic factor neurturin (NTN) and mediates activation of the RET tyrosine kinase receptor, which is essential for downstream neurotrophic signaling.
Neuronal Development: It is required for the development of the myenteric nerve plexus and parasympathetic islet innervation, making it critical for studies on the enteric nervous system and disorders such as Hirschsprung disease.
Disease Modeling: GFRα-2 is implicated in the survival and maintenance of specific neuronal populations, including isolectin B4-binding nociceptive sensory neurons, and is thus relevant for research on neuropathies, neurodegeneration, and pain mechanisms.
Therapeutic Screening: Recombinant GFRα-2 can be used in cell-based assays to study ligand-receptor interactions, screen for agonists/antagonists, and evaluate neuroprotective compounds targeting the GDNF family signaling pathways.
Biochemical and Structural Studies: The recombinant protein enables in vitro biochemical assays, binding studies, and structural analyses to elucidate the molecular mechanisms of GFRα-2-mediated signaling.
High Purity and Defined Activity: Recombinant preparations offer high purity and batch-to-batch consistency, which are essential for reproducible experimental results in cell culture, ELISA, and other in vitro applications.
Typical applications include:
Neuronal survival and differentiation assays
RET receptor activation studies
Disease modeling for enteric neuropathies and neurodegenerative disorders
Screening of neurotrophic factors and small molecules
ELISA standards and positive controls in immunoassays
In summary, recombinant Mouse GFRα-2 is a valuable tool for dissecting neurotrophic signaling pathways, modeling disease, and developing therapeutic strategies targeting the GDNF family receptor system.
You can use recombinant Mouse GFRα-2 as a standard for quantification or calibration in your ELISA assays, provided that the recombinant protein is of high purity, its concentration is accurately determined, and it is compatible with your assay’s antibodies and detection system.
Key considerations:
Purity and Quantification: The recombinant protein should be highly purified and its concentration precisely measured, ideally by absorbance at 280 nm or another validated method.
Source and Sequence: The recombinant GFRα-2 should match the native protein’s sequence and post-translational modifications as closely as possible, especially if your ELISA detects conformational epitopes or glycosylation-dependent sites.
Validation: It is essential to validate that the recombinant standard produces a dose-response curve parallel to that of the endogenous protein in your sample matrix. This ensures accurate quantification and minimizes matrix effects.
Standard Curve Preparation: Prepare the standard curve using serial dilutions of the recombinant protein, following best practices for ELISA standard preparation.
Documentation: Refer to the product’s datasheet or certificate of analysis for specific instructions on reconstitution, dilution, and compatibility with your assay format.
Additional notes:
Some commercial ELISA kits for mouse GFRα-2 use recombinant protein as the standard, confirming its suitability for this purpose.
If your ELISA is custom-developed, ensure that the recombinant standard is recognized equivalently to the native protein by your capture and detection antibodies.
Always run pilot experiments to confirm linearity, parallelism, and recovery when introducing a new standard.
In summary, recombinant Mouse GFRα-2 is appropriate for use as a standard in ELISA quantification, provided you validate its performance in your specific assay context.
Recombinant Mouse GFRα-2 has been validated for several applications in published research, primarily in studies of neurobiology, receptor-ligand interactions, and disease models involving the GDNF (glial cell line-derived neurotrophic factor) family signaling pathways.
Key validated applications include:
In vitro receptor-ligand binding assays: Recombinant Mouse GFRα-2 is used to study binding interactions with its ligand, neurturin (NTN), and to characterize the activation of downstream signaling pathways, such as RET tyrosine kinase activation.
ELISA (Enzyme-Linked Immunosorbent Assay): The protein is used as a standard or capture reagent in direct ELISAs to quantify or detect GFRα-2 or its ligands in biological samples.
Western blotting: Recombinant Mouse GFRα-2 is used as a positive control or to validate antibody specificity in Western blot assays, confirming the detection of GFRα-2 in tissue or cell lysates.
Immunohistochemistry (IHC): Antibodies validated against recombinant GFRα-2 have been used for IHC to localize GFRα-2 expression in tissue sections, particularly in studies of the nervous system and pancreas.
Functional cell-based assays: Recombinant GFRα-2 is used to reconstitute or modulate GDNF family signaling in cultured cells, enabling the study of downstream effects such as neuronal survival, differentiation, or hormone secretion.
Disease model research: The protein has been used in studies investigating its role in the development of the myenteric nerve plexus (relevant to Hirschsprung disease), pancreatic innervation, and neuropathies associated with chronic pancreatitis.
Supporting details:
Recombinant Mouse GFRα-2 is frequently used as a tool to validate antibody specificity in both Western blot and ELISA formats, as indicated by multiple antibody datasheets and product descriptions.
Published studies have used recombinant GFRα-2 to investigate its role in neuronal development and disease, including its necessity for the development of the myenteric plexus and its involvement in pancreatic neuropathy.
Immunohistochemistry validation is supported by manufacturer data and published research, where recombinant protein is used to confirm antibody reactivity and tissue localization.
Summary Table of Validated Applications
Application
Description/Context
Receptor-ligand binding
In vitro assays for GDNF/NTN binding and RET activation
ELISA
Standard or capture reagent for quantification/detection
Western blot
Positive control or antibody validation
Immunohistochemistry (IHC)
Antibody validation and tissue localization studies
Functional cell assays
Reconstitution of signaling, neuronal survival/differentiation studies
Disease model research
Studies of enteric nervous system, pancreatic innervation, neuropathies, HSCR
If you require protocols or more specific examples from primary literature, please specify the application of interest.
To reconstitute and prepare Recombinant Mouse GFRα-2 protein for cell culture experiments, use sterile technique and follow these steps:
Centrifuge the vial briefly (20–30 seconds in a microcentrifuge) before opening to ensure all lyophilized powder is at the bottom.
Reconstitute the protein in sterile, endotoxin-free phosphate-buffered saline (PBS), pH 7.2–7.3, or sterile distilled water if PBS is not specified for your application.
Recommended concentration for reconstitution is typically between 0.1–1.0 mg/mL. For example, add 100–1000 µL of buffer to 100 µg of protein to achieve 1 mg/mL to 0.1 mg/mL, respectively.
Gently mix by pipetting up and down. Do not vortex, as this may denature the protein.
Allow the protein to dissolve completely at room temperature for several minutes. Inspect visually to confirm dissolution.
Aliquot the solution to avoid repeated freeze-thaw cycles, which can degrade the protein.
Storage after reconstitution:
Short-term: 2–8°C for up to one month.
Long-term: ≤–20°C (preferably –70°C) for up to 3–12 months.
Avoid repeated freeze-thaw cycles.
Additional notes for cell culture use:
Confirm protein integrity by SDS-PAGE if needed.
For sensitive cell types, consider adding carrier protein (e.g., 0.1% BSA) to minimize adsorption and loss, unless experimental design requires otherwise.
Ensure endotoxin levels are suitable for cell culture (<1 EU/µg is typical for recombinant GFRα-2).
Dilute the reconstituted stock into cell culture medium immediately before use to achieve the desired final concentration.
Products are for research use only. Not for use in diagnostic or therapeutic procedures.
