Recombinant Human CNTF Rα

Recombinant Human CNTF Rα is used in research applications to study and modulate the signaling of ciliary neurotrophic factor (CNTF), particularly in contexts involving neuronal survival, differentiation, and neuroprotection. It serves as the ligand-specific α-receptor for CNTF, enabling precise investigation of CNTF-mediated pathways and cellular responses.

Key scientific reasons to use Recombinant Human CNTF Rα:

• Mechanistic Studies: CNTF Rα is essential for reconstituting the functional CNTF receptor complex in vitro, which consists of CNTF Rα, gp130, and LIFRβ. This allows researchers to dissect the molecular mechanisms of CNTF signaling, including downstream effects on neuronal and glial cells.
• Cellular Responsiveness: Recombinant CNTF Rα can be used to confer CNTF responsiveness to cells that naturally lack the receptor, enabling controlled studies of CNTF’s effects on diverse cell types. Soluble CNTF Rα (sCNTF Rα) can render cells expressing only gp130 and LIFRβ responsive to CNTF, broadening experimental possibilities.
• Neuroprotection and Regeneration: CNTF signaling via CNTF Rα has demonstrated neuroprotective effects in models of retinal degeneration, optic nerve injury, and neurodegenerative diseases. Using recombinant CNTF Rα allows for targeted activation or inhibition of these pathways to study therapeutic potential.
• Protein Interaction and Binding Assays: Recombinant CNTF Rα is valuable for biochemical assays, such as ligand binding, receptor activation, and protein-protein interaction studies, facilitating the characterization of CNTF and its receptor complex.
• Drug Screening and Development: Recombinant receptor proteins are critical tools for screening CNTF agonists, antagonists, or modulators, which may have therapeutic applications in neurodegenerative or inflammatory diseases.

Best practices for research applications:

• Use recombinant CNTF Rα in combination with CNTF and other receptor components to reconstitute signaling in cell-based assays.
• Employ recombinant CNTF Rα to study species-specific differences in receptor expression and function, which is important for translational research.
• Validate the biological activity and stability of recombinant CNTF Rα under experimental conditions to ensure reproducibility and reliability of results.

Summary:
Recombinant Human CNTF Rα is a critical reagent for investigating CNTF signaling, enabling mechanistic, therapeutic, and translational research in neurobiology and regenerative medicine. Its use allows for precise control and modulation of CNTF pathways, supporting studies on neuronal survival, differentiation, and neuroprotection.

Using Recombinant Human CNTF Rα as an ELISA Standard

No, recombinant human CNTF Rα is not appropriate for use as a standard for quantification or calibration in ELISA assays. CNTF Rα is a receptor component, not the ligand itself, and serves a fundamentally different role in the CNTF signaling pathway.

Key Distinctions

CNTF Rα Function: CNTF Rα is the alpha receptor subunit that binds ciliary neurotrophic factor (CNTF). It is part of the receptor complex required for CNTF signaling, not a quantifiable analyte in standard ELISA protocols designed to measure CNTF levels.

Appropriate Standards: For ELISA assays measuring CNTF, you should use recombinant human CNTF protein itself as your standard. Recombinant CNTF protein is specifically designed and validated for this purpose, with well-characterized biological activity. For example, recombinant human CNTF demonstrates an ED₅₀ of 0.03-0.18 μg/mL for stimulating cell proliferation in the TF-1 human erythroleukemic cell line.

Proper ELISA Design Considerations

When establishing CNTF quantification assays, use purified recombinant CNTF protein to prepare your standard curve. The recombinant protein should be reconstituted according to specifications—typically at 100 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin.

If your research specifically requires measuring CNTF Rα levels rather than CNTF itself, dedicated CNTF Rα ELISA kits are available with appropriate recombinant CNTF Rα standards formulated for that specific application.

Recombinant Human CNTF Receptor Alpha (CNTF Rα) has been validated for several key applications in published research, primarily in studies of neuroprotection, cell signaling, and neuronal differentiation.

Validated Applications:

• Cell-based bioassays: CNTF Rα is used to assess the biological activity of CNTF and its ability to mediate cell survival and proliferation, often in combination with recombinant human CNTF. Soluble CNTF Rα (sCNTFRα) can combine with CNTF to mediate cell survival, demonstrating ligand specificity and affinity comparable to the membrane-bound receptor.

• Neuroprotection studies: CNTF Rα is central to research on neurodegenerative diseases, such as retinitis pigmentosa and amyotrophic lateral sclerosis, where it mediates the neuroprotective effects of CNTF on retinal cells and neurons. Studies have shown that CNTF Rα is involved in promoting the survival of retinal ganglion cells and photoreceptors, both in vitro and in vivo.

• Neuronal differentiation and stem cell research: CNTF Rα is used in protocols for neural stem cell proliferation, differentiation of induced pluripotent stem cell (iPSC)-derived neural progenitors, Schwann cells, and astrocytes, as well as in the culture of retinal cells. It is also applied to induce axonal growth and study neural stem cell biology.

• Signal transduction analysis: CNTF Rα is employed to study downstream signaling pathways, such as STAT3 phosphorylation, in various cell types including preadipocyte 3T3-L1 cells. This helps elucidate the molecular mechanisms of CNTF-mediated effects.

• Disease modeling: CNTF Rα is used in animal models and ex vivo retinal explant cultures to investigate the penetration, distribution, and efficacy of CNTF in treating retinal degeneration and other neurological disorders.

Additional Notes:

• CNTF Rα is often studied in both its membrane-bound and soluble forms, as the latter can confer CNTF responsiveness to cells lacking the membrane-bound receptor.
• The receptor is validated for use with human, mouse, rat, and other species in bioassays.
• Its role in mediating the effects of modified CNTF molecules with enhanced potency and stability has also been explored in therapeutic research.

In summary, Recombinant Human CNTF Rα is validated for use in bioassays, neuroprotection studies, neuronal differentiation protocols, signal transduction analysis, and disease modeling, with a strong focus on its role in mediating CNTF’s effects on neuronal survival and differentiation.

To reconstitute and prepare Recombinant Human CNTF Rα protein (Ciliary Neurotrophic Factor Receptor Alpha) for cell culture experiments, follow these best-practice steps:

• Centrifuge the vial briefly before opening to ensure all lyophilized protein is at the bottom.
• Reconstitute the protein using a suitable buffer. For CNTF Rα, a common choice is 5–10 mM sodium phosphate, pH 7.5, or sterile PBS, pH 7.4. If the product datasheet specifies a different buffer, follow that recommendation.
• Add sterile water or buffer to achieve a final concentration of 0.1–1.0 mg/mL. For example, for 100 μg of protein, add 100–1000 μL of buffer.
• Gently pipette the solution up and down to dissolve the protein. Avoid vortexing or vigorous pipetting, as this can denature the protein.
• If the protein does not dissolve completely or a precipitate forms, centrifuge the solution and use only the clear supernatant for experiments.
• For long-term storage, aliquot the reconstituted protein and store at –20°C or –80°C. Avoid repeated freeze-thaw cycles.
• For short-term use, the reconstituted protein can be stored at 4°C for up to 2–7 days.
• To enhance stability, especially for dilute solutions or long-term storage, add a carrier protein such as 0.1–1% BSA or HSA. This is particularly important if the protein will be stored at low concentrations or subjected to freeze-thaw cycles.

Summary protocol:

1. Centrifuge vial.
2. Add sterile buffer (e.g., 5–10 mM sodium phosphate, pH 7.5, or PBS, pH 7.4) to reach 0.1–1.0 mg/mL.
3. Gently pipette to dissolve; do not vortex.
4. If needed, centrifuge to remove insoluble material.
5. Aliquot and store at –20°C or –80°C with carrier protein if desired.

Application note:
Before use in cell culture, dilute the reconstituted stock to the desired working concentration using cell culture medium. If using serum-free medium, ensure the presence of a carrier protein to minimize adsorption losses.

Always consult the specific product datasheet for any unique requirements or recommendations for your particular recombinant CNTF Rα preparation.