Recombinant Rat IL-1β

Recombinant Rat IL-1β is widely used in research applications because it is a key proinflammatory cytokine that enables precise modeling and manipulation of inflammatory, immune, and cell signaling pathways in rat-based experimental systems.

Essential context and supporting details:

• Inflammation and Immunology Studies: IL-1β is central to the regulation of inflammation, cell proliferation, and apoptosis. Using recombinant rat IL-1β allows researchers to study these processes in vitro and in vivo, particularly in rat models relevant to autoimmune diseases, neuroinflammation, and metabolic disorders.

• Disease Modeling: Recombinant rat IL-1β is used to induce or modulate disease states such as arthritis, encephalomyelitis, diabetes, and wound healing in rat models, enabling the investigation of pathophysiological mechanisms and therapeutic interventions.

• Cellular and Molecular Mechanisms: It is employed to stimulate specific cell types (e.g., T cells, beta cells, neurons) to analyze downstream signaling events, gene expression changes, and cytokine interactions. For example, IL-1β enhances NMDA receptor-mediated neuronal responses and can be used to study neurotoxicity and neuroprotection.

• Bioassays and Functional Studies: Recombinant rat IL-1β is used in bioassays to assess cell proliferation, cytokine secretion, and functional responses in rat cells, providing a standardized and reproducible stimulus for experimental protocols.

• Translational Research: It supports drug development and biomarker discovery by serving as a target or stimulus in preclinical studies, including testing IL-1β inhibitors or receptor antagonists for anti-inflammatory therapies.

• Tumor Biology: IL-1β facilitates tumor progression, metastasis, and angiogenesis in animal models, making it valuable for cancer research and immuno-oncology applications.

Best practices:

• Use recombinant rat IL-1β at defined concentrations to ensure reproducibility and relevance to physiological or pathological conditions.
• Select appropriate assay formats (e.g., ELISA, bioassay, in vivo administration) based on experimental goals.
• Confirm species specificity to avoid cross-reactivity and ensure accurate modeling of rat biology.

Summary of scientific applications:

• Modeling inflammation and immune responses in rats
• Studying cytokine signaling and cell-cell interactions
• Testing anti-inflammatory drugs and biologics
• Investigating mechanisms of disease progression and tissue repair
• Analyzing tumor microenvironment and cancer metastasis

Using recombinant rat IL-1β provides a controlled, species-specific tool for dissecting complex biological processes and validating therapeutic strategies in rat-based research systems.

Yes, recombinant rat IL-1β can be used as a standard for quantification or calibration in ELISA assays, provided it is compatible with your assay system and matches the form of IL-1β detected by your antibodies. Recombinant standards are commonly used in commercial rat IL-1β ELISA kits and in custom assays, but there are important considerations for accurate quantification.

Key points and supporting details:

• Recombinant rat IL-1β is widely used as a standard in ELISA kits. Many commercial ELISA kits for rat IL-1β use recombinant protein as the calibrator to generate the standard curve, allowing quantification of both recombinant and endogenous IL-1β in samples.

• Assay compatibility is essential. The recombinant standard should be of the same form (e.g., mature vs. precursor) as the IL-1β detected by your assay antibodies. Some ELISA kits are calibrated using mature IL-1β, and these may underestimate precursor forms. Check whether your antibodies recognize the same isoform as your standard.

• Parallelism and validation. For accurate quantification, the recombinant standard must behave in a parallel manner to the endogenous protein in your sample matrix. This is typically validated by the kit manufacturer, but if you are developing your own assay, you should confirm parallelism between the standard curve and serially diluted biological samples.

• Reconstitution and diluent. Follow the recommended reconstitution protocol and use the same diluent as specified for the standard curve in your ELISA protocol to avoid matrix effects.

• Carrier protein and formulation. Some recombinant proteins are supplied with carrier proteins (e.g., BSA) for stability, which is generally acceptable for ELISA standards, but ensure this does not interfere with your assay.

• Documentation and traceability. Use a recombinant standard with a well-documented sequence, purity, and activity, ideally with a certificate of analysis, to ensure reproducibility and traceability in your quantification.

In summary:
You can use recombinant rat IL-1β as a standard for ELISA quantification if it is validated for your assay system, matches the detected isoform, and is prepared according to best practices for ELISA calibration. Always confirm compatibility with your specific antibodies and assay conditions.

Recombinant Rat IL-1β has been validated for a diverse range of applications across multiple research domains, as demonstrated in published literature.

Cell-Based Bioassays

The protein has been extensively used in bioassay applications to assess cellular responses. It stimulates cell proliferation of the D10.G4.1 mouse helper T cell line in a dose-dependent manner, with established potency parameters. This bioassay application has been documented across numerous publications, making it a standard tool for evaluating IL-1β biological activity.

In Vivo Disease Models

Recombinant Rat IL-1β has been applied in several in vivo experimental models to study disease mechanisms and therapeutic interventions:

• Autoimmune and Neuroinflammatory Models: The protein has been used to investigate experimental autoimmune encephalomyelitis (EAE) severity and disease progression, particularly in studies examining regulatory immune cell activity.

• Retinal Degeneration: Intravitreal administration of recombinant IL-1β has been employed to promote oxidative stress and apoptosis in retinal tissue, serving as a model for studying diabetic retinopathy and retinal neovascularization.

• Nociceptive Processing: Spinal application of IL-1β in anesthetized rats has been used to examine mechanical hyperexcitability and spinal cord neuron responses to stimulation, with findings indicating microglia and IL-6 signaling involvement.

• Wound Healing and Chemotherapy Effects: The protein has been utilized in studies examining IL-1β-mediated signaling in chemotherapy-induced alopecia and wound healing responses.

Immunological and Inflammatory Studies

The protein serves as a tool for investigating IL-1β receptor interactions and signaling pathways. Its species-specific activity makes it particularly valuable for rat studies, as human and mouse IL-1β variants bind poorly to rat IL-1 receptors.

Structural and Functional Characterization

Recombinant Rat IL-1β has been validated for analytical applications including SDS-PAGE, HPLC, mass spectrometry, and functional studies to confirm protein integrity and biological activity.

To reconstitute and prepare Recombinant Rat IL-1β protein for cell culture experiments, follow these steps based on best practices and multiple technical datasheets:

1. Centrifuge the vial briefly before opening to ensure all lyophilized protein is at the bottom.

2. Reconstitution buffer selection:

   • For most cell culture applications, use sterile phosphate-buffered saline (PBS), pH 7.2–7.4.
   • If the datasheet specifies, you may also use sterile distilled water. However, PBS is generally preferred for stability and physiological compatibility.

3. Protein concentration:

   • Common reconstitution concentrations are 100 μg/mL in sterile PBS or 0.1–0.5 mg/mL in sterile distilled water.
   • If using for sensitive cell culture work, add 0.1% carrier protein (such as bovine serum albumin, BSA) to the PBS to minimize adsorption and loss of activity.

4. Reconstitution procedure:

   • Add the calculated volume of buffer directly to the vial to achieve the desired concentration.
   • Gently swirl or invert the vial; do not vortex. Allow the protein to dissolve completely (typically 10–30 minutes at room temperature).
   • If any undissolved material remains, gently pipette up and down or let stand longer.

5. Aliquoting and storage:

   • Once fully dissolved, aliquot the solution into small volumes to avoid repeated freeze-thaw cycles.
   • Store aliquots at –20°C to –70°C for long-term storage. For short-term use (up to 1 month), store at 2–8°C under sterile conditions.
   • Avoid repeated freeze-thaw cycles, as this can reduce protein activity.

6. Working solution preparation:

   • Before adding to cell cultures, dilute the stock solution to the desired working concentration using sterile cell culture medium or PBS with 0.1% BSA.
   • Typical working concentrations for bioassays are in the ng/mL range (e.g., ED₅₀ for D10.G4.1 cell proliferation is 0.3–2.7 ng/mL).

Summary of key steps:

• Use sterile PBS (with 0.1% BSA if possible) for reconstitution.
• Reconstitute to 100 μg/mL (or as required).
• Gently dissolve, aliquot, and store at –20°C or below.
• Dilute to working concentration in cell culture medium before use.

Additional notes:

• Always consult the specific product datasheet for any unique instructions, as formulations may vary.
• If using for in vivo or highly sensitive assays, ensure all reagents are endotoxin-free.

These steps will ensure optimal solubility, stability, and biological activity of recombinant rat IL-1β for cell culture experiments.