Recombinant Human IL-1α

Recombinant Human IL-1α is a valuable tool for a wide range of research applications due to its central role in inflammation, immune regulation, and tissue homeostasis. Here are several compelling reasons to use Recombinant Human IL-1α in your research:

1. Key Mediator of Inflammation

IL-1α is a potent pro-inflammatory cytokine that initiates and amplifies inflammatory responses. It acts as an "alarmin," rapidly released from damaged or dying cells to alert the immune system during sterile injury, infection, or tissue stress. This makes it essential for studying:

• Sterile inflammation (e.g., ischemia, trauma)
• Innate immune activation
• Early immune responses to tissue damage

2. Broad Biological Activity

Recombinant IL-1α is biologically active and can stimulate a variety of cellular responses in vitro and in vivo, including:

• Cell proliferation (e.g., T cells, fibroblasts, vascular smooth muscle cells)
• Induction of other cytokines and chemokines (e.g., IL-6, IL-8, TNFα)
• Production of matrix metalloproteinases (MMPs) and other enzymes involved in tissue remodeling
• Activation of osteoclasts and chondrocytes, making it relevant for bone and cartilage research

3. Use as a Positive Control

IL-1α is widely used as a positive control in assays involving inflammation, immune cell activation, and tissue remodeling. For example:

• In chondral explant studies, IL-1α reliably induces markers of osteoarthritis (e.g., COMP, MMP13).
• In immune cell assays, it activates T cells, monocytes, and macrophages.

4. Role in Disease Models

IL-1α is implicated in the pathogenesis of several diseases, including:

• Autoinflammatory syndromes
• Rheumatoid arthritis
• Cancer (angiogenesis, tumor invasiveness, cachexia)
• Aging-related inflammation

Using recombinant IL-1α allows researchers to model these conditions and test potential therapeutic interventions.

5. High Specificity and Purity

Recombinant IL-1α is available in highly purified, carrier-free forms, ensuring minimal batch-to-batch variability and reducing the risk of confounding effects from contaminants.

6. Compatibility with Diverse Assays

Recombinant IL-1α is suitable for a wide range of experimental applications, including:

• In vitro bioassays (cell proliferation, cytokine production)
• Western blotting and other protein detection methods
• In vivo studies (animal models of inflammation, cancer, or tissue injury)

7. Therapeutic Target Validation

IL-1α is a validated target for therapeutic intervention. Using recombinant IL-1α in your research allows you to:

• Study the effects of IL-1α blockade (e.g., with neutralizing antibodies or receptor antagonists)
• Validate the efficacy of anti-inflammatory drugs

8. Relevance to Translational Research

Findings using recombinant IL-1α can be directly translated to clinical settings, as IL-1α inhibitors are already in use or under investigation for treating inflammatory diseases, cancer, and autoimmune disorders.

In summary, Recombinant Human IL-1α is a versatile and reliable reagent for studying inflammation, immune responses, and disease mechanisms. Its broad biological activity, high purity, and relevance to both basic and translational research make it an essential tool for any laboratory investigating inflammatory or immune-related processes.

Yes, recombinant human IL-1α can be used as a standard for quantification or calibration in ELISA assays, provided it is compatible with your assay system. This is a common practice in research ELISAs for cytokines such as IL-1α.

Key considerations and supporting details:

• Recombinant human IL-1α is widely used as an ELISA standard: Many commercial ELISA kits for human IL-1α use recombinant protein as the calibrator to generate the standard curve. The standard is typically reconstituted and serially diluted to cover the assay’s dynamic range, allowing for accurate quantification of IL-1α in samples.

• Compatibility with your assay: The recombinant IL-1α standard should match the form (e.g., mature, precursor, or fragment) recognized by the capture and detection antibodies in your ELISA. Most kits specify that their antibodies detect both natural and recombinant forms of IL-1α, but you should confirm this in your assay documentation or by consulting the antibody datasheets.

• Standard preparation: Follow the manufacturer’s instructions for reconstitution and dilution of the recombinant IL-1α standard. Serial dilutions are typically prepared directly in the assay plate or in tubes to generate a standard curve covering the expected concentration range.

• Validation: If you are developing a custom ELISA or using a kit with different antibodies, validate that your recombinant IL-1α standard produces a linear, parallel standard curve and that sample quantification is accurate. Parallelism between the standard curve and sample dilution curves is a key indicator of assay validity.

• Source and purity: Ensure the recombinant IL-1α is of high purity and free from contaminants that could interfere with the assay. The protein should be stored and handled according to the supplier’s recommendations to maintain stability and activity.

Summary Table: Use of Recombinant Human IL-1α as ELISA Standard

In summary: Recombinant human IL-1α is suitable and commonly used as a standard in ELISA assays for quantification and calibration, provided it is compatible with your assay’s antibodies and protocol. Always validate performance in your specific assay context.

Recombinant Human IL-1α has been validated for a wide range of applications in published research, including:

• Bioassay: Used to assess biological activity in various cell types, including whole cells and chondral explants, often as a positive control for inflammatory responses.
• Cell Culture: Employed to stimulate cell proliferation (e.g., T cells, fibroblasts), induce collagenase and prostaglandin production, and study inflammatory signaling pathways.
• Control: Serves as a control in experimental setups to induce or modulate inflammatory responses.
• ELISA (Standard): Used as a standard in ELISA assays to quantify IL-1α levels.
• In Vivo: Applied in animal models to study inflammatory diseases, immune responses, and therapeutic interventions.
• Western Blot: Utilized to detect and analyze IL-1α protein expression and its effects on signaling pathways.
• Cell Migration/Motility: Investigated for its role in recruiting infiltrating cells to sites of injury and inflammation.
• Sterile Inflammation Studies: Used to study the initiation of sterile inflammation, particularly in processes involving necrosis and alarmin release.

These applications highlight the versatility of Recombinant Human IL-1α in both basic research and translational studies, particularly in the context of inflammation, immune response, and disease modeling.

To reconstitute and prepare Recombinant Human IL-1α protein for cell culture experiments, dissolve the lyophilized protein in sterile PBS (pH 7.4), ideally supplemented with 0.1% endotoxin-free human or bovine serum albumin (HSA or BSA) to a working concentration suitable for your assay, commonly 10–200 μg/mL depending on experimental needs.

Step-by-step protocol:

• Briefly centrifuge the vial before opening to ensure all lyophilized material is at the bottom.
• Add sterile PBS (pH 7.4), optionally containing 0.1% HSA or BSA to minimize adsorption and stabilize the protein.
• Do not vortex; gently swirl or tap the vial to mix until fully dissolved.
• Typical reconstitution concentrations range from 10 μg/mL (for bioassays) up to 0.2 mg/mL (for stock solutions).
• After reconstitution, aliquot the solution to avoid repeated freeze-thaw cycles.
• Store aliquots at 2–8°C for up to 1 month or at –20°C for long-term storage.
• For cell culture, further dilute the reconstituted stock in your culture medium to the desired final concentration, ensuring the buffer is compatible with your cells.

Additional notes:

• If the product datasheet recommends a different buffer (e.g., sterile water), follow those instructions, but PBS with carrier protein is standard for cytokines like IL-1α.
• Always use endotoxin-free reagents to prevent unwanted immune activation in cell culture.
• Avoid vigorous mixing or vortexing, which can denature the protein.
• For sensitive applications, filter the final solution through a 0.2 μm sterile filter if sterility is a concern.

Summary Table:

This protocol ensures optimal solubility, stability, and biological activity of recombinant human IL-1α for cell culture experiments.