Recombinant Mouse Eotaxin

Recombinant Mouse Eotaxin (CCL11) is widely used in research applications to study immune cell migration, allergic inflammation, and eosinophil biology due to its role as a potent and selective chemoattractant for eosinophils.

Key scientific reasons to use recombinant mouse eotaxin in your research:

• Eosinophil Chemotaxis: Eotaxin specifically induces migration of eosinophils, making it essential for investigating mechanisms of eosinophil recruitment in models of asthma, allergy, and parasitic infection.
• Immune Response Regulation: Eotaxin binds to the CCR3 receptor on eosinophils, facilitating their accumulation at sites of inflammation and contributing to the study of Th2-driven immune responses.
• Bioassays and Functional Studies: Recombinant eotaxin is validated for use in bioactivity assays, enabling precise quantification of eosinophil chemotaxis, calcium flux, and cell signaling in vitro and in vivo.
• ELISA Standard: It serves as a reference standard for ELISA, allowing accurate measurement of eotaxin levels in biological samples.
• Pathway Analysis: Eotaxin participates in key signaling pathways (MAPK/ERK, NF-κB), making it useful for dissecting molecular mechanisms of cell migration, proliferation, and inflammation.
• Modeling Disease Mechanisms: Recombinant mouse eotaxin is critical for modeling allergic airway inflammation, neuroinflammation, and tissue eosinophilia in mouse models, supporting translational research in immunology and respiratory disease.

Additional technical advantages:

• High Purity and Activity: Recombinant preparations are typically >95% pure and endotoxin-free, ensuring reproducibility and minimizing confounding effects in sensitive assays.
• Defined Sequence and Tag-Free Options: Recombinant proteins provide consistent activity and are available in tag-free formats, which is important for functional and structural studies.
• Compatibility with Multiple Applications: Suitable for use in cell culture, tissue models, in vivo studies, and analytical techniques such as SDS-PAGE, HPLC, and mass spectrometry.

In summary, recombinant mouse eotaxin is an essential reagent for dissecting eosinophil biology, allergic inflammation, and immune signaling in mouse models, offering high specificity, reproducibility, and versatility for a wide range of experimental protocols.

Yes, recombinant mouse Eotaxin (CCL11) is routinely used as a standard for quantification or calibration in ELISA assays designed to measure mouse Eotaxin levels in biological samples.

Context and Supporting Details:

• ELISA kits for mouse Eotaxin/CCL11 typically include a lyophilized recombinant mouse Eotaxin protein as the standard. This standard is reconstituted and serially diluted to generate a calibration curve, which is then used to quantify Eotaxin concentrations in unknown samples.
• The recombinant standard is specifically validated for use in ELISA and is intended to provide accurate, reproducible quantification of both recombinant and endogenous (natural) mouse Eotaxin in matrices such as serum, plasma, and cell culture supernatants.
• Protocols for these ELISA kits specify the use of recombinant mouse Eotaxin as the standard, and the assay is optimized to ensure that the recombinant protein is recognized equivalently to the native protein in biological samples.

Best Practices:

• Ensure that the recombinant Eotaxin standard you use is of high purity and has been validated for ELISA applications. Most commercial ELISA kits provide a recombinant standard that is quality-controlled for this purpose.
• Follow the kit or assay protocol for reconstitution, dilution, and handling of the recombinant standard to maintain accuracy and reproducibility.
• If you are developing your own ELISA (not using a commercial kit), confirm that your antibodies recognize both recombinant and native Eotaxin equivalently, as minor differences in folding or post-translational modifications can sometimes affect antibody binding.

Limitations:

• Recombinant proteins used as standards are typically produced in E. coli or other expression systems and may lack certain post-translational modifications present in native proteins. However, for most ELISA applications, this does not significantly affect quantification, as the antibodies are selected to recognize epitopes present on both forms.
• Always verify that the recombinant standard is suitable for your specific ELISA format and detection antibodies, especially if you are assembling a custom assay.

Summary Table: Use of Recombinant Mouse Eotaxin as ELISA Standard

Conclusion:
You can use recombinant mouse Eotaxin as a standard for quantification or calibration in ELISA assays, provided it is validated for this purpose and used according to established protocols.

Recombinant Mouse Eotaxin (CCL11) has been validated for several key applications in published research, including bioassays, ELISA (as a standard), Western blot, and in vivo studies.

Validated Applications:

• Bioassay:
  Used to assess chemotactic activity, particularly for eosinophils, and to study cellular responses such as calcium flux, eosinophil survival, and basophil histamine release.
  Frequently applied in studies of inflammatory responses, allergic disease models, and immune cell migration.

• ELISA (Standard):
  Employed as a reference standard for quantifying eotaxin levels in biological samples, including serum, plasma, and cell culture supernatants.
  Used in research on asthma, allergic inflammation, and other eosinophil-associated conditions.

• Western Blot:
  Utilized for protein detection and quantification in tissue or cell lysates, validating eotaxin expression in various experimental contexts.

• In Vivo Studies:
  Applied in mouse models to investigate the physiological and pathological roles of eotaxin, such as its involvement in allergic airway inflammation, neurogenesis suppression during aging, and inflammatory bone resorption.
  Used to study eosinophil recruitment, immune cell trafficking, and disease mechanisms in live animals.

Additional Context:

• Cell and Tissue Culture:
  Recombinant eotaxin is recommended for use in cell and tissue culture experiments, often with BSA as a carrier to enhance stability.

• Disease Models:
  Extensively used in models of asthma, allergic rhinitis, atopic dermatitis, helminth infection, and inflammatory bone diseases.
  Also implicated in studies of aging, neurogenesis, and cognitive decline.

• Mechanistic Studies:
  Validated for investigating signaling pathways (e.g., NF-κB, STAT6), cellular senescence, and oxidative stress responses in vitro.

Summary Table of Validated Applications

Note:
Most published research utilizes recombinant mouse eotaxin for functional studies of eosinophil biology, allergic inflammation, and as a biomarker in immunological assays. It is also a standard reagent for validating disease mechanisms and therapeutic interventions in preclinical models.

To reconstitute and prepare Recombinant Mouse Eotaxin (CCL11) protein for cell culture experiments, dissolve the lyophilized protein at 100 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin (BSA). This approach helps stabilize the protein and prevent adsorption to plastic surfaces, which is critical for reproducible cell culture applications.

Detailed protocol:

• Centrifuge the vial briefly before opening to ensure all lyophilized material is at the bottom.
• Add sterile PBS (phosphate-buffered saline) to achieve a final concentration of 100 μg/mL. For example, add 100 μL PBS to 100 μg protein.
• Include at least 0.1% BSA in the PBS if the protein is not already supplied with carrier protein. This is especially important for cell culture use to minimize protein loss due to adsorption.
• Gently mix by pipetting up and down or by gentle vortexing. Avoid vigorous agitation to prevent denaturation.
• Allow the protein to dissolve at room temperature for 10–30 minutes, ensuring complete reconstitution.
• Aliquot the solution into single-use volumes to avoid repeated freeze-thaw cycles, which can degrade the protein.
• Storage after reconstitution:
  • Store aliquots at 2–8 °C for up to 1 month.
  • For longer-term storage, keep at –20 °C to –70 °C.
  • Avoid repeated freeze-thaw cycles.

Additional notes:

• If the product is supplied carrier-free, the addition of BSA or another carrier protein is strongly recommended for cell culture applications.
• If the protein is supplied with BSA as a carrier, reconstitution in PBS alone is sufficient.
• For dilution to working concentrations (e.g., 1–100 ng/mL for cell stimulation), use sterile PBS with 0.1% BSA as the diluent to maintain protein stability.
• Always consult the product-specific datasheet or certificate of analysis for any unique instructions, as formulations may vary between suppliers.

Summary table:

This protocol ensures optimal stability and biological activity of recombinant mouse Eotaxin for cell culture experiments.