Recombinant Human IL-1β

Recombinant Human IL-1β is widely used in research because it is a potent pro-inflammatory cytokine that plays a central role in inflammation, immune regulation, and disease pathogenesis, making it essential for studying cellular responses, disease mechanisms, and therapeutic interventions.

Key reasons to use Recombinant Human IL-1β in research applications:

• Modeling Inflammatory Responses: IL-1β is a primary mediator of inflammation and is crucial for host defense against infection and injury. Adding recombinant IL-1β to cell cultures or animal models allows researchers to mimic inflammatory conditions and study downstream effects, such as cytokine production (e.g., IL-6), cell proliferation, and apoptosis.

• Mechanistic Studies: IL-1β is central to inflammasome activation and signaling through the IL-1 receptor (IL-1R1), which triggers a cascade of inflammatory mediators. Recombinant IL-1β enables precise dissection of these pathways in vitro and in vivo.

• Drug Discovery and Validation: Recombinant IL-1β is used to screen and validate anti-inflammatory drugs, including IL-1β inhibitors and monoclonal antibodies, by providing a reproducible stimulus for pharmacodynamic and mechanistic studies.

• Immunology and Cell Biology: It is used to stimulate immune cells (e.g., T cells, B cells, macrophages) to study differentiation, proliferation, and cytokine secretion, as well as to investigate the role of IL-1β in autoimmune and autoinflammatory diseases.

• Disease Modeling: IL-1β is implicated in diseases such as rheumatoid arthritis, gout, type 2 diabetes, and neuroinflammation. Recombinant IL-1β is used to induce disease-like states in experimental models, facilitating the study of pathogenesis and therapeutic intervention.

• Biomarker and Assay Development: Recombinant IL-1β serves as a standard or positive control in assays such as ELISA, flow cytometry, and bioassays for quantifying cytokine responses and validating antibody specificity.

• Reproducibility and Consistency: Recombinant proteins provide high purity, defined activity, and batch-to-batch consistency, which are critical for generating reliable and interpretable experimental data.

In summary, Recombinant Human IL-1β is indispensable for research on inflammation, immune signaling, and therapeutic development due to its well-characterized biological activity and central role in disease-relevant pathways.

Yes, recombinant human IL-1β can be used as a standard for quantification or calibration in ELISA assays, provided it is compatible with the specific ELISA kit and detection system you are using.

Most commercial ELISA kits for human IL-1β are calibrated using recombinant human IL-1β as the reference standard. For example:

• Many kits (such as those from R&D Systems, BioLegend, Abcam, Medix Biochemica, and others) explicitly state that their assays are calibrated against recombinant human IL-1β, and the standard provided in the kit is typically recombinant IL-1β.
• The recombinant IL-1β standard is used to generate a standard curve, which allows for the quantification of IL-1β in your samples.
• It is important to reconstitute and dilute the recombinant IL-1β according to the manufacturer’s instructions, and to use a fresh standard for each assay.

However, keep in mind:

• The form of IL-1β (mature vs. precursor) may affect detection. Most ELISA kits are calibrated with the mature form, and may not accurately detect or quantify the unprocessed precursor.
• If you are using a third-party recombinant IL-1β (not from the kit manufacturer), ensure it is of high purity and has been validated for use in your specific ELISA system.

In summary, recombinant human IL-1β is suitable for use as a standard in ELISA assays for quantification and calibration, but always follow the protocol and recommendations for your specific kit and application.

Recombinant Human IL-1β has been validated for a broad range of applications in published research, primarily as a bioactive cytokine for cell-based assays, immunological studies, and as a positive control in detection and quantification assays.

Key validated applications include:

• Bioassays: Used to stimulate cell proliferation (e.g., D10.G4.1 mouse helper T cell line), induce cytokine release (such as IL-6 from human fibroblasts), and model inflammatory responses in vitro.
• Cell Culture: Applied to study immune cell activation, differentiation (e.g., Th17 cells), and B cell proliferation.
• Screening Assays: Serves as a target for screening inhibitory molecules, including monoclonal antibodies (e.g., canakinumab) and small-molecule antagonists that block IL-1β signaling.
• Detection and Quantification: Used as a positive control in ELISA, sELISA, and other immunoassays to quantify IL-1β levels.
• Functional Studies: Investigates the role of IL-1β in inflammation, immune modulation, and disease models such as rheumatoid arthritis, Crohn’s disease, osteoarthritis, and neuroinflammatory conditions.
• In Vivo Models: Validated for use in animal studies to assess immune responses, inflammation, and therapeutic interventions targeting IL-1β signaling.
• Protein Characterization: Utilized in SDS-PAGE, HPLC, mass spectrometry, and other biochemical assays for protein analysis.

Additional validated uses in published research:

• Mechanistic Studies: Elucidation of IL-1β’s role in signaling pathways (e.g., MAPK, NF-κB), apoptosis, and cellular metabolism.
• Therapeutic Research: Evaluation of IL-1β antagonists and neutralizing antibodies in models of autoinflammatory and autoimmune diseases.
• Immunomodulation: Studies on activation of B and T cells, regulatory T cell function, and innate lymphoid cell responses.

These applications are supported by extensive literature and product validation data, demonstrating the versatility of recombinant human IL-1β in immunology, inflammation, and therapeutic research.

To reconstitute and prepare Recombinant Human IL-1β protein for cell culture experiments, dissolve the lyophilized protein in sterile water or buffer to a concentration between 0.1–1.0 mg/mL. For optimal stability and activity in cell culture, further dilute the stock solution in sterile PBS or culture medium containing a carrier protein such as 0.1–1% BSA or HSA to prevent adsorption and loss of activity.

Step-by-step protocol:

• Bring vial contents to the bottom by brief centrifugation before opening.
• Add sterile water or buffer (e.g., PBS) to achieve a concentration of 0.1–1.0 mg/mL. For example, add 100–1000 μL to 100 μg of protein.
• Gently mix by pipetting or inversion; avoid vigorous vortexing to prevent protein denaturation.
• If not used immediately, aliquot the stock solution and store at –20 °C or below to avoid repeated freeze-thaw cycles.
• For working solutions, dilute the stock in cell culture medium or PBS containing 0.1–1% BSA or HSA to the desired final concentration (e.g., 1–100 ng/mL for most cell assays).
• Filter sterilize if necessary, using a 0.2 μm filter, especially if reconstituted in non-sterile conditions.

Additional recommendations:

• Use carrier protein (BSA or HSA) in all dilutions to minimize loss due to adsorption to plasticware.
• Avoid repeated freeze-thaw cycles by aliquoting stock solutions.
• For sensitive applications, confirm endotoxin levels are suitable for cell culture (typically <1 EU/μg).

Example calculation:
To reconstitute 10 μg of IL-1β to 0.1 mg/mL, add 100 μL of sterile water or PBS. For a 1 mg/mL stock, add 10 μL.

Summary Table:

These guidelines ensure maximal activity and stability of recombinant IL-1β for cell culture experiments.