Recombinant Mouse IL-1β

Recombinant Mouse IL-1β is a valuable tool for research applications due to its central role in inflammation, immune regulation, and host defense. Here are several key reasons why you should consider using recombinant Mouse IL-1β in your studies:

1. Mimics Physiological IL-1β Activity

Recombinant Mouse IL-1β closely replicates the biological activity of endogenous IL-1β, allowing researchers to study its effects in a controlled and reproducible manner. It activates key signaling pathways such as NF-κB, JNK, and p38 MAPK, which are involved in the expression of inflammatory cytokines and chemokines.

2. Stimulates Immune and Inflammatory Responses

IL-1β is a potent proinflammatory cytokine that:

• Triggers the production of other proinflammatory cytokines (e.g., IL-6, IL-8).
• Promotes the extravasation of immunocompetent cells into affected tissues.
• Stimulates Th17 differentiation and B cell proliferation.
• Initiates acute-phase responses to infection and injury.

3. Supports Diverse Experimental Applications

Recombinant Mouse IL-1β is suitable for a wide range of research applications, including:

• Cell culture studies to investigate immune cell activation and differentiation.
• Bioassays to measure cytokine activity and cellular responses.
• In vivo assays to study the effects of IL-1β on disease models such as sepsis, autoimmune disorders, and inflammation.
• ELISA standards and Western Blot controls for quantifying and detecting IL-1β in experimental samples.

4. Enhances Innate Immunity and Host Defense

Studies have shown that recombinant IL-1β can:

• Improve survival in sepsis models by stimulating local proliferation and differentiation of bone marrow cells into regulatory dendritic cells.
• Enhance bacterial eradication and alleviate sepsis by activating innate immune responses.
• Protect against experimental sepsis and other inflammatory conditions.

5. High Bioactivity and Purity

Recombinant Mouse IL-1β produced in systems like Pichia pastoris (yeast) offers natural folding and post-translational modifications, resulting in superior bioactivity compared to proteins derived from E. coli. These proteins are typically free from endotoxins, HIS-tags, and carrier proteins, ensuring consistent and reliable results.

6. Facilitates Mechanistic Studies

Using recombinant IL-1β allows researchers to dissect the molecular mechanisms underlying IL-1β signaling, including its role in:

• Inflammation and immune regulation.
• Cell proliferation, differentiation, and survival.
• Disease pathogenesis and therapeutic intervention.

7. Versatile for Different Research Models

Recombinant Mouse IL-1β is compatible with various experimental models, including:

• Whole cell cultures.
• Transgenic and knockout mice.
• In vivo disease models.

Summary

Recombinant Mouse IL-1β is essential for studying inflammation, immune responses, and host defense mechanisms. Its high bioactivity, purity, and versatility make it an ideal choice for a wide range of research applications, from basic mechanistic studies to translational research in disease models.

Yes, recombinant Mouse IL-1β can be used as a standard for quantification or calibration in ELISA assays, provided it is properly validated for your specific assay system.

Recombinant Mouse IL-1β is widely employed as a quantitative standard in sandwich ELISA formats to measure mouse IL-1β protein levels. Commercial ELISA kits routinely use recombinant Mouse IL-1β to generate standard curves, enabling accurate quantification of IL-1β in biological samples such as cell culture supernatants, tissue lysates, serum, and plasma. The standard curve is typically prepared using serial dilutions of the recombinant protein, and sample concentrations are interpolated from this curve.

Key considerations for use:

• Validation: Ensure the recombinant standard is compatible with the antibodies and detection system in your ELISA. Most kits are calibrated against recombinant Mouse IL-1β, and dose-response curves for natural and recombinant forms are generally parallel, indicating equivalence for quantification.
• Reconstitution and Dilution: Follow the manufacturer’s instructions for reconstitution and dilution of the recombinant standard to maintain accuracy and reproducibility.
• Dynamic Range: Prepare a standard curve covering the expected concentration range in your samples (e.g., 12.5–2000 pg/mL).
• Specificity: Confirm that your assay specifically detects Mouse IL-1β and does not cross-react with related cytokines or proteins.

Limitations:

• Use recombinant Mouse IL-1β standards only for research purposes, not for diagnostic procedures, unless specifically validated for clinical use.
• The accuracy of quantification depends on proper standard curve preparation and assay optimization.

Best Practices:

• Always run a fresh standard curve with each assay to ensure quantitative reliability.
• Use the same diluent for standards and samples to avoid matrix effects.
• Store recombinant standards as recommended to preserve activity and stability.

In summary, recombinant Mouse IL-1β is suitable and commonly used as a standard for ELISA quantification, provided assay-specific validation and best practices are followed.

Recombinant Mouse IL-1β has been validated for a wide range of applications in published research, primarily focusing on its role as a pro-inflammatory cytokine in immunological, neuroinflammatory, and disease models. Key applications include:

• Bioassays and Functional Assays: Used to stimulate immune and inflammatory responses in vitro, such as measuring proliferation of immune cells (e.g., D10S cells), cytokine production, and activation of signaling pathways (e.g., AKT/mTOR, SIRT1/ERK, inflammasome activation).
• Cell Culture Studies: Employed to study the effects of IL-1β on cell differentiation, activation, and function, including microglia, astrocytes, chondrocytes, and bone mesenchymal stem cells.
• In Vivo Assays: Administered in animal models to induce inflammation, study disease progression (e.g., osteoarthritis, septic endoplasmic reticulum stress, traumatic brain injury), and evaluate therapeutic interventions.
• ELISA Standards: Used as a standard in sandwich ELISA to quantify IL-1β levels in biological samples.
• Western Blot Controls: Serves as a positive control in Western blotting to confirm antibody specificity and protein expression.
• Neuroinflammatory Disease Models: Investigated in models of neuroinflammation, including Alzheimer’s disease, multiple sclerosis, and diabetic retinopathy, to understand the role of IL-1β in neuronal injury and disease progression.
• Cardiovascular and Autoimmune Disease Models: Utilized in studies of cardiovascular complications and autoimmune diseases to assess the impact of IL-1β inhibition and its therapeutic potential.

These applications highlight the versatility of recombinant Mouse IL-1β in both basic and translational research, providing insights into the mechanisms of inflammation and potential therapeutic targets.

To reconstitute and prepare Recombinant Mouse IL-1β protein for cell culture experiments, dissolve the lyophilized protein in sterile buffer (typically water or PBS) containing a carrier protein such as BSA or FCS, aiming for a final concentration between 0.1–1.0 mg/mL. Use low endotoxin reagents and handle under sterile conditions.

Step-by-step protocol:

1. Centrifuge the vial briefly before opening to ensure all lyophilized material is at the bottom.
2. Add sterile buffer:
   • Use sterile distilled water or phosphate-buffered saline (PBS) as the solvent.
   • Include at least 0.1% carrier protein (e.g., BSA or heat-inactivated FCS) to stabilize the protein and prevent adsorption to plastic.
   • For example, to reconstitute 100 µg of protein, add 100 µL (for 1 mg/mL) to 1 mL (for 0.1 mg/mL) of buffer.
3. Gently mix the solution by pipetting up and down or swirling; do not vortex.
4. Aliquot the reconstituted protein into small volumes to avoid repeated freeze-thaw cycles.
5. Storage:
   • Store aliquots at –20°C or colder for long-term use.
   • For short-term use (up to 1 week), store at 2–8°C.
6. Working dilutions:
   • Prepare further dilutions in cell culture medium or buffer containing carrier protein immediately before use.
   • Use low endotoxin medium for sensitive cell types.

Additional notes:

• If the protein appears as a film, ensure complete dissolution by gentle mixing.
• Avoid repeated freeze-thaw cycles to maintain protein activity.
• Always consult the specific product datasheet for any unique instructions.

Summary of key points:

• Buffer: Sterile water or PBS with ≥0.1% BSA or FCS.
• Concentration: 0.1–1.0 mg/mL for stock.
• Mixing: Gentle, no vortexing.
• Aliquot and store: –20°C or colder.
• Working dilutions: Prepare fresh in medium with carrier protein.

This protocol ensures optimal stability and biological activity of recombinant mouse IL-1β for cell culture applications.