Recombinant Human Artemin

Recombinant Human Artemin (ARTN) is a valuable tool for research applications due to its well-characterized biological functions and its role in multiple signaling pathways relevant to neuroscience, development, and disease. Here are several key reasons why you should consider using Recombinant Human Artemin in your research:

1. Neuroprotection and Neuronal Survival

Artemin is a member of the glial cell line-derived neurotrophic factor (GDNF) family and is known to promote the survival of various neuronal populations, including:

• Sensory and sympathetic neurons
• Dopaminergic midbrain neurons
• Peripheral ganglia neurons

This makes it highly useful for studies on neuroprotection, neurodegeneration, and neuronal regeneration.

2. Neurite Outgrowth and Differentiation

Artemin supports neurite outgrowth and differentiation of neurons, making it ideal for:

• Studies on neural development
• Investigation of axon guidance and regeneration
• Screening for compounds or conditions that enhance neuronal growth

3. Modulation of Pain and Neuropathic Conditions

Artemin has been shown to mitigate neuropathic pain and reverse morphological changes associated with nerve injury. It is relevant for:

• Research on pain mechanisms
• Development of therapies for neuropathic pain
• Preclinical models of nerve injury and repair

4. Role in Cancer and Disease

Emerging evidence suggests Artemin is involved in tumorigenesis, metastasis, and therapeutic resistance in certain cancers (e.g., endometrial carcinoma, mammary tumors). This makes it useful for:

• Cancer biology studies
• Investigation of tumor microenvironment and signaling pathways
• Development of targeted therapies

5. Well-Defined Molecular and Functional Properties

Recombinant Human Artemin is typically produced as a non-glycosylated, disulfide-linked homodimer with high purity and reproducible bioactivity. It is suitable for:

• Functional assays (e.g., ELISA, cell-based assays)
• In vivo and in vitro studies
• Standardization of experimental conditions

6. Receptor and Signaling Pathway Studies

Artemin signals through the GFRα3/RET receptor complex and has been shown to interact with NCAM (neural cell adhesion molecule), providing opportunities to:

• Study complex signaling mechanisms
• Investigate receptor-ligand interactions
• Develop mimetics or inhibitors

7. Therapeutic Potential

Artemin and its mimetics (e.g., artefin) have shown promise in preclinical and clinical trials for neurological disorders and pain management, supporting translational research.

In summary, Recombinant Human Artemin is a versatile and biologically active protein that can advance research in neuroscience, neuroprotection, pain, cancer, and regenerative medicine. Its well-documented effects and availability in high-purity, bioactive forms make it a reliable choice for a wide range of experimental applications.

Yes, recombinant human Artemin can be used as a standard for quantification or calibration in ELISA assays, provided it is of high purity and properly reconstituted. This approach is commonly used when a purified native protein standard is unavailable.

Essential context and supporting details:

• Recombinant Protein as ELISA Standard: Using a recombinant protein as an ELISA standard is a standard practice, especially when the native protein is difficult to obtain or purify. The recombinant human Artemin should be well-characterized, and its concentration accurately determined, ideally by an independent method such as HPLC or UV absorbance.
• Formulation Considerations: Recombinant Artemin is available in formulations with or without carrier proteins (e.g., BSA). For ELISA standards, the version with BSA is generally recommended to enhance stability and reproducibility, unless BSA interferes with your assay.
• Reconstitution: Follow the manufacturer’s instructions for reconstitution, as improper handling can affect protein stability and quantification accuracy.
• Calibration and Lot-to-Lot Variation: When using recombinant proteins as standards, be aware that the immunologically recognizable mass may differ from the mass stated on the vial due to post-translational modifications or folding differences. It is best practice to assign the standard’s value based on its performance in the ELISA, rather than relying solely on the vial label.
• Validation: Validate the standard curve generated with recombinant Artemin by running known concentrations and assessing recovery, linearity, and precision. This ensures the standard is suitable for your specific ELISA system.

Additional relevant information:

• ELISA Kits: Commercial Artemin ELISA kits typically include a recombinant Artemin standard for calibration, demonstrating its suitability for this purpose.
• Best Practices: Prepare fresh standards for each assay, use appropriate diluents, and avoid repeated freeze-thaw cycles to maintain protein integrity.
• Limitations: If your ELISA is highly sensitive to protein conformation or glycosylation, ensure the recombinant Artemin closely mimics the native protein’s structure. Some assays may show slight discrepancies in quantification due to these factors.

In summary, recombinant human Artemin is appropriate for use as an ELISA standard, but careful validation and handling are required for accurate quantification.

Recombinant Human Artemin has been validated for several key applications in published research, primarily in the fields of neurobiology, cancer biology, and cell signaling.

Validated Applications:

• Bioassays (Functional and Cell-Based):

  • Used to assess neuronal survival, neurite outgrowth, and neuroprotection in primary neuronal cultures and cell lines.
  • Demonstrated to induce phosphorylation of Akt (PKB) in SH-SY5Y neuroblastoma cells, indicating activation of intracellular signaling pathways.
  • Shown to promote sensory and sympathetic neuron survival and support dopaminergic midbrain neurons in vitro.

• In Vivo Studies:

  • Applied in animal models of nerve injury to evaluate its effects on neuronal regeneration, neuropathic pain relief, and neurochemical normalization.
  • Used in orthotopic transplantation models to study its role in tumor invasiveness and perineural invasion in pancreatic adenocarcinoma.

• ELISA (Enzyme-Linked Immunosorbent Assay):

  • Employed as a capture reagent in ELISA to study ligand-receptor interactions, such as binding to GFRα3/RET and NCAM.

• Immunohistochemistry (IHC):

  • Utilized to detect Artemin expression in tissue microarrays of human tumors, particularly pancreatic adenocarcinoma, to correlate expression with clinical features.

• Surface Plasmon Resonance (SPR):

  • Used to characterize protein-protein interactions, such as binding to its receptors or other ligands.

Additional Context and Supporting Details:

• Neurobiology: Artemin is a member of the GDNF family of ligands and is widely used to study neurotrophic effects, including neuronal survival, axonal growth, and regeneration after injury.
• Cancer Research: Artemin has been implicated in tumor progression, invasiveness, and perineural invasion in several cancers, including pancreatic, breast, and lung cancers. Functional studies often use recombinant Artemin to modulate these processes in vitro and in vivo.
• Signaling Pathways: Recombinant Artemin is used to activate and study downstream signaling pathways, such as RET tyrosine kinase and Akt phosphorylation, in various cell types.
• Therapeutic Potential: Preclinical and clinical studies have evaluated recombinant Artemin for neuropathic pain relief and neurodegenerative disease models, supporting its translational relevance.

Summary Table of Validated Applications

These applications are well-supported by published research and demonstrate the versatility of recombinant human Artemin in both basic and translational studies.

To reconstitute and prepare Recombinant Human Artemin protein for cell culture experiments, dissolve the lyophilized protein at 100 μg/mL in sterile 4 mM HCl containing at least 0.1% human or bovine serum albumin (BSA). If your formulation does not contain BSA, you may reconstitute in sterile 4 mM HCl alone. Alternatively, some protocols allow reconstitution in sterile water at 0.1 mg/mL (100 μg/mL), but the use of acid and carrier protein is preferred for stability and activity.

Step-by-step protocol:

• Centrifuge the vial briefly before opening to collect all lyophilized material at the bottom.
• Add sterile 4 mM HCl (with 0.1% BSA if recommended) to achieve a final concentration of 100 μg/mL.
• Gently pipette up and down and wash down the sides of the vial to ensure complete dissolution.
• Allow the protein to dissolve for several minutes at room temperature, avoiding vigorous mixing or vortexing.
• Once fully dissolved, the solution can be further diluted in cell culture medium or other aqueous buffers as required for your experiment.

Storage and handling:

• After reconstitution, store the solution at 4–8 °C for up to 2–7 days.
• For longer-term storage, aliquot and freeze at <–20 °C to avoid repeated freeze-thaw cycles.
• Always use a manual defrost freezer and avoid repeated freeze-thaw cycles to preserve protein activity.

Additional notes:

• The recommended working concentration for bioassays is typically in the range of 4–16 ng/mL, but optimize based on your cell type and experimental design.
• Ensure all solutions and diluents are sterile and endotoxin-free to prevent adverse effects on cell cultures.
• If using for sensitive neuronal cultures, confirm the absence of contaminants and verify protein activity in a pilot assay.

Summary Table:

Always consult the specific product datasheet for any additional manufacturer recommendations regarding reconstitution and handling.