Recombinant Mouse IL-1α

Recombinant Mouse IL-1α is a valuable tool for research applications due to its critical roles in immune regulation and inflammatory responses. Here are the key reasons to incorporate it into your research:

Immune and Inflammatory Functions

IL-1α is a central cytokine in both innate and adaptive immune responses. It functions as a key regulator of inflammatory processes and plays a determinative role in amplifying cytokine responses initiated by innate immune stimuli. The cytokine is expressed by multiple cell types including epithelial cells, activated macrophages, neutrophils, and endothelial cells, making it highly relevant for studying diverse biological systems.

Specific Biological Activities

When using the recombinant form, you gain access to a protein with high specific biological activity suitable for in vitro assays. The protein demonstrates potent bioactivity, with effective concentrations in the picogram-per-milliliter range—recombinant mouse IL-1α stimulates cell proliferation of helper T cell lines with an ED₅₀ of 3-7 pg/mL.

Research Applications

T cell activation and proliferation: IL-1α induces T cell activation during antigen presentation and autonomous proliferation in various cell types.

Immune cell regulation: The cytokine stimulates monocyte/macrophage-mediated tumor cytotoxicity and promotes osteoclast formation.

Chemokine expression: IL-1α induces expression of chemokine mRNA and proteins, such as Cxcl1, in specialized cell populations like mouse granulosa cells.

Tissue responses: IL-1α recruits infiltrating cells to sites of injury during necrosis and contributes to angiogenesis during hypoxia.

Systemic effects: Intranasal administration of recombinant IL-1α induces robust pro-inflammatory responses and generates transcriptomic signatures comparable to viral infection models.

Experimental Advantages

Using recombinant IL-1α allows you to establish dose-dependent responses in controlled experimental settings, study specific signaling pathways independent of complex cellular contexts, and model inflammatory conditions relevant to immune dysregulation, autoimmune diseases, and infectious disease responses.

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

Key considerations and supporting details:

• Scientific Basis: ELISA kits for mouse IL-1α routinely use recombinant mouse IL-1α as the standard to generate calibration curves for quantification. These standards are typically produced in E. coli or mammalian expression systems and are validated to ensure that the dose-response curves for recombinant and native IL-1α are parallel, allowing accurate quantification of both forms in biological samples.

• Validation: It is essential that the recombinant IL-1α you use as a standard is of high purity and bioactivity, and that its sequence and post-translational modifications (if any) are compatible with the antibodies used in your ELISA. Commercial ELISA kits often provide their own recombinant standard, which has been validated for use with their specific antibody pairs and assay conditions.

• Assay Compatibility: If you are using a commercial ELISA kit, it is strongly recommended to use the standard provided with the kit, as it has been specifically validated for that assay. Substituting with a different recombinant IL-1α may introduce variability due to differences in protein folding, glycosylation, or other modifications, potentially affecting antibody recognition and quantification accuracy.

• Custom or In-House ELISA: If you are developing your own ELISA or using a custom assay, you can use recombinant mouse IL-1α as a standard, but you must validate that:

  • The standard curve generated with your recombinant protein is linear and reproducible within the assay’s dynamic range.
  • The recombinant standard produces a dose-response curve parallel to that of native IL-1α in your sample matrix.
  • There is no significant cross-reactivity or interference with other proteins present in your samples.

• Documentation: Always document the source, lot number, and concentration of the recombinant IL-1α used, and include validation data in your experimental records.

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

In summary: You can use recombinant mouse IL-1α as a standard for ELISA quantification if it is validated for your assay system, and it is best practice to use the standard provided with commercial kits unless you have performed appropriate validation experiments.

Recombinant Mouse IL-1α has been validated for several key applications in published research, primarily in bioassays, in vitro cell culture, in vivo studies, and as an ELISA standard.

Validated Applications:

• Bioassays: Used to stimulate cell proliferation (e.g., D10.G4.1 mouse helper T cell line), assess cytokine activity, and study immune cell activation and differentiation.
• In Vitro Cell Culture: Applied to mouse granulosa cells to induce chemokine expression (e.g., CXCL1), vascular smooth muscle cell proliferation, T cell activation, monocyte/macrophage-mediated cytotoxicity, and osteoclast formation.
• In Vivo Studies: Employed in mouse models to investigate inflammatory responses, immune modulation, neurodegeneration, and disease mechanisms such as autoimmune encephalomyelitis, liver injury, and respiratory viral infection.
• ELISA Standard: Used as a quantitative standard in ELISA assays for cytokine measurement.
• Cell Migration/Motility Assays: Validated for studies on cell migration and motility, particularly in the context of inflammation and tissue injury.
• Stem/Immune Cell Maintenance or Differentiation: Utilized to study the effects on stem and immune cell populations.

Representative Published Research Applications:

• Neuroinflammation and Neurodegeneration: Investigating microglial inflammatory responses and tauopathy in mouse models.
• Tumor Immunology: Studying IL-1α secretion in oral squamous cell carcinoma and its impact on immune suppression.
• Autoimmunity: Exploring the role of IL-1α in experimental autoimmune encephalomyelitis and central nervous system inflammation.
• Renal Inflammation: Demonstrating IL-1α-dependent inflammation in diabetic nephropathy models.
• Respiratory Viral Infection: Assessing the role of IL-1α in weight loss and immune response following RSV infection in mice.
• Cellular Differentiation and Activation: Examining keratinocyte differentiation, osteoclastogenesis, and T cell activation.

Additional Notes:

• In vivo administration of recombinant IL-1α (e.g., intranasal delivery) has been shown to induce acute inflammatory responses, weight loss, and upregulation of other cytokines in mouse models.
• In vitro studies often use recombinant IL-1α to dissect signaling pathways, cytokine networks, and cellular responses relevant to both innate and adaptive immunity.

These applications are supported by multiple peer-reviewed studies and product validation data, confirming the utility of recombinant Mouse IL-1α in diverse immunological, inflammatory, and cell biology research contexts.

To reconstitute and prepare Recombinant Mouse IL-1α protein for cell culture experiments, follow these general guidelines based on manufacturer recommendations and best practices:

Reconstitution

1. Centrifuge the vial: Before opening, centrifuge the lyophilized protein vial briefly (20–30 seconds in a microcentrifuge) to ensure all powder is at the bottom.

2. Reconstitution buffer: Most recombinant mouse IL-1α proteins are reconstituted in sterile water or sterile PBS. Some protocols recommend adding 0.1% BSA (bovine serum albumin) to stabilize the protein, especially if you plan to store aliquots.

   • Typical reconstitution concentration: 0.1 mg/mL (100 µg/mL) in sterile water or PBS, sometimes with 0.1% BSA.
   • Example: For a 10 µg vial, add 100 µL of sterile water or PBS (with or without 0.1% BSA).

3. Gentle mixing: After adding the buffer, gently pipette up and down or swirl the vial to dissolve the protein. Avoid vortexing or vigorous shaking to prevent foaming and protein denaturation.

Preparation for Cell Culture

1. Aliquot and store: If not using immediately, aliquot the reconstituted protein into small volumes and store at ≤ –20°C (preferably –80°C) to avoid repeated freeze-thaw cycles.

2. Dilution: For cell culture experiments, dilute the reconstituted IL-1α in your cell culture medium or an appropriate buffer to the desired working concentration. Typical working concentrations for bioassays range from 1–100 ng/mL, but optimal concentration should be determined empirically for your specific cell type and assay.

3. Handling: Always thaw frozen aliquots on ice and use immediately after thawing. Avoid repeated freeze-thaw cycles.

Additional Tips

• Sterility: Maintain sterile conditions throughout the process to prevent contamination.
• Avoid foaming: When mixing or diluting, avoid foaming as it can denature the protein.
• Check manufacturer’s instructions: Always refer to the specific product datasheet or certificate of analysis for any unique requirements.

Example Protocol

1. Centrifuge the vial for 20–30 seconds.
2. Add 100 µL sterile water (or PBS with 0.1% BSA) to a 10 µg vial.
3. Gently mix until fully dissolved.
4. Aliquot into sterile tubes and store at –80°C.
5. For cell culture, dilute to working concentration in culture medium and add to cells.

By following these steps, you can ensure the stability and activity of recombinant mouse IL-1α for your cell culture experiments.