Recombinant Rat IL-1α

Recombinant Rat IL-1α is used in research to model and dissect the biological roles of interleukin-1 alpha, a key pro-inflammatory cytokine, in rat systems. It is particularly valuable for studying inflammation, immune responses, and disease mechanisms relevant to both basic and translational research.

Key reasons to use recombinant rat IL-1α in research applications:

• Mimics Endogenous Cytokine Activity: Recombinant rat IL-1α closely replicates the activity of the natural cytokine, allowing for controlled studies of its effects on target cells and tissues in vitro and in vivo.
• Inflammation and Immune Response Studies: IL-1α is a potent mediator of inflammation, promoting the release of other inflammatory mediators, acute phase proteins, and supporting hematopoietic stem cell expansion. It is widely used to induce and study inflammatory pathways, immune cell activation, and cytokine signaling in rat models.
• Disease Modeling: Administration of recombinant IL-1α can be used to model diseases such as rheumatoid arthritis, neuroinflammation, and fever, as it triggers hallmark symptoms like joint inflammation, fever, and activation of the hypothalamic-pituitary-adrenal axis. This is critical for preclinical testing of anti-inflammatory drugs and understanding disease mechanisms.
• Reproducibility and Standardization: Recombinant proteins provide consistent, defined activity and purity, which is essential for reproducible experimental results and for quantifying biological responses.
• Assay Development and Validation: Recombinant rat IL-1α is used as a standard in ELISA and other immunoassays to quantify endogenous IL-1α levels in biological samples, ensuring assay accuracy and comparability.

Additional context:

• Mechanistic Studies: By adding recombinant IL-1α to cell cultures or animal models, researchers can dissect downstream signaling pathways, gene expression changes, and cellular responses specific to IL-1α stimulation.
• Comparative Studies: Using recombinant rat IL-1α allows for direct comparison with other cytokines (such as IL-1β) or with antagonists (such as IL-1ra), helping to clarify the distinct and overlapping roles of these molecules in inflammation and immunity.

In summary, recombinant rat IL-1α is a critical tool for investigating the molecular and cellular mechanisms of inflammation, validating therapeutic targets, and developing new interventions in rat models.

Yes, you can use Recombinant Rat IL-1α as a standard for quantification or calibration in your ELISA assays, provided that the recombinant protein is highly purified and its concentration is accurately determined. The use of recombinant cytokines as standards is a common and accepted practice in immunoassays, including ELISA, as long as the recombinant standard is recognized by the antibodies used in your assay and behaves similarly to the native protein in terms of immunoreactivity.

Several commercial ELISA kits for Rat IL-1α (such as the Quantikine Rat IL-1 alpha ELISA kit and others referenced in your search results) are calibrated using E. coli-expressed recombinant Rat IL-1α. These kits demonstrate that recombinant IL-1α can be used to generate reliable standard curves for quantifying both recombinant and native IL-1α in various sample types.

Key Points to Consider:

• Purity and Concentration: Ensure the recombinant IL-1α is of high purity and that its concentration is accurately known (e.g., determined by amino acid analysis or other validated methods).
• Immunoreactivity: Confirm that the recombinant protein is recognized by the capture and detection antibodies in your ELISA system.
• Matrix Effects: Dilute the standard in a matrix similar to your samples (e.g., assay buffer or sample diluent) to minimize matrix effects.
• Parallelism: Ideally, the dose-response curve of the recombinant standard should be parallel to that of the native IL-1α in your samples.

References:

• R&D Systems Quantikine Rat IL-1 alpha ELISA kit uses recombinant Rat IL-1α as the standard and is calibrated against it.
• Many other ELISA kits for Rat IL-1α (e.g., ELK Biotech, Elabscience, RayBiotech) also use recombinant standards for calibration.

In summary, recombinant Rat IL-1α is suitable for use as a standard in ELISA assays for quantification, as long as it is properly characterized and validated for your specific assay conditions.

Recombinant Rat IL-1α has been validated primarily for use in SDS-PAGE and is widely used as a standard or control in biochemical assays, but published research does not indicate extensive validation for functional cell-based assays or in vivo models.

Key validated applications in published research and technical documentation:

• SDS-PAGE: The most consistently validated application is SDS-PAGE, where recombinant rat IL-1α is used to confirm purity, molecular weight, and integrity of the protein preparation. This is a standard quality control step for recombinant proteins.
• Biochemical and immunological assays: Recombinant rat IL-1α is described as suitable for use in immunology and inflammation studies, suggesting its use as a cytokine standard or positive control in ELISA, Western blot, or related assays. However, explicit published validation for these applications in peer-reviewed research is not detailed in the available sources.
• Research use only: Most sources specify that the protein is for research use only, with no direct evidence of validation in functional cell-based assays or animal models in the rat system.

Additional context:

• While recombinant IL-1α from other species (such as human) has been used in functional studies (e.g., stimulating cytokine release, cell proliferation, or in vivo inflammation models), there is no direct evidence in the provided search results that recombinant rat IL-1α has been validated for these applications in published research.
• Some vendors mention its use in immunology and inflammation studies, which implies potential for cell signaling or cytokine induction assays, but without explicit peer-reviewed validation data.

Summary Table:

If you require information on a specific functional assay (e.g., cytokine induction, cell proliferation), please specify, as current published research does not provide direct validation for these applications with recombinant rat IL-1α.

To reconstitute and prepare Recombinant Rat IL-1α protein for cell culture experiments, follow these steps for optimal activity and stability:

1. Centrifuge the vial briefly before opening to ensure all lyophilized protein is collected at the bottom.
2. Reconstitution:
   • Add sterile distilled water to achieve a final concentration of 0.1–0.5 mg/mL.
   • Alternatively, you may use a sterile aqueous buffer (e.g., PBS, pH 7.4) if required for your application.
   • Avoid vortexing or vigorous pipetting to prevent protein denaturation.
3. Carrier Protein/Stabilizer (recommended for long-term storage or dilute solutions):
   • Add a carrier protein such as 0.1% BSA, 5% HSA, 10% FBS, or 5% trehalose to stabilize the protein and minimize adsorption to tube walls.
4. Aliquoting:
   • Divide the reconstituted solution into small aliquots to avoid repeated freeze-thaw cycles, which can degrade the protein.
5. Storage:
   • Store aliquots at -20°C for up to 3 months.
   • For short-term use (up to 1 week), store at 2–8°C.
   • The lyophilized protein is stable at -20°C to -80°C for up to 1 year.
6. Preparation for Cell Culture:
   • Before adding to cell culture, dilute the reconstituted stock to the desired working concentration using sterile culture medium or buffer.
   • Ensure the final buffer conditions are compatible with your cell type (e.g., isotonicity, pH).

Additional Notes:

• The protein is typically supplied lyophilized from PBS, pH 7.4, and is of high purity (≥90% by SDS-PAGE) with low endotoxin (<1 EU/μg).
• Avoid repeated freeze-thaw cycles and vigorous mixing to preserve biological activity.
• If using for sensitive cell types, confirm endotoxin levels are suitable for your application.

Summary Table: Recombinant Rat IL-1α Reconstitution

These steps ensure the protein remains stable and active for cell culture experiments.