Recombinant Human IFN-alpha 2b

Recombinant Human IFN-alpha 2b is widely used in research due to its potent antiviral, antiproliferative, and immunomodulatory properties, making it a valuable tool for studying immune responses, cancer biology, and viral pathogenesis.

Key scientific reasons to use recombinant human IFN-alpha 2b in research applications:

• Antiviral Activity: IFN-alpha 2b is a type I interferon that induces the expression of interferon-stimulated genes (ISGs), which inhibit viral replication and spread. It is commonly used to model and investigate antiviral mechanisms in cell culture and animal models.
• Antiproliferative and Antitumor Effects: IFN-alpha 2b suppresses cell proliferation and induces apoptosis in various cancer cell lines. It is FDA-approved for the treatment of several malignancies, including hairy cell leukemia, malignant melanoma, and Kaposi's sarcoma, and is used in research to study cancer cell biology and immune-mediated tumor suppression.
• Immunomodulation: IFN-alpha 2b modulates innate and adaptive immune responses by activating cytotoxic T cells and natural killer (NK) cells, and by influencing cytokine production. This makes it useful for dissecting immune signaling pathways and studying immunotherapy strategies.
• Standardization and Reproducibility: Recombinant production ensures high purity, batch-to-batch consistency, and defined biological activity, which are critical for reproducible experimental results and reliable bioassays.
• Clinical Relevance: IFN-alpha 2b is the representative type I IFN subtype in both basic and translational research, reflecting its widespread clinical use in antiviral and antineoplastic therapies. Its mechanisms—primarily via the JAK-STAT pathway—are well-characterized, facilitating mechanistic studies.
• Versatility: It is used in diverse applications, including:
  • Viral infection models
  • Cancer cell proliferation and apoptosis assays
  • Immune cell activation and cytokine profiling
  • Combination therapy studies with other drugs

In summary, recombinant human IFN-alpha 2b is a scientifically validated cytokine for research in virology, oncology, and immunology, offering robust, reproducible, and clinically relevant experimental outcomes.

Yes, recombinant human IFN-alpha 2b can be used as a standard for quantification or calibration in ELISA assays, provided it is of high purity and its concentration is accurately determined. This practice is well-established in research and clinical assay development for interferon quantification.

Key considerations and best practices:

• Purity and Activity: Ensure the recombinant IFN-alpha 2b is highly purified and biologically active. Most commercial preparations specify purity (≥95%) and provide bioactivity data, which is essential for reliable calibration.
• Carrier Protein: Recombinant IFN-alpha 2b is available in carrier-free and BSA-containing formulations. For ELISA standards, BSA-containing formulations are often recommended to enhance stability and prevent adsorption to plasticware, unless BSA interferes with your assay.
• Standard Curve Preparation: Accurately reconstitute and dilute the recombinant protein using the same buffer or diluent as your samples to minimize matrix effects. Serial dilutions should cover the expected range of IFN-alpha concentrations in your samples.
• Validation: Confirm that your ELISA detects both recombinant and native IFN-alpha 2b equivalently. Most commercial ELISA kits are validated for both forms, but it is good practice to check recovery and linearity in your specific sample matrix.
• Documentation: Record the lot number, concentration, and storage conditions of your standard. Some protocols recommend aliquoting and freezing standards to avoid repeated freeze-thaw cycles.

Scientific precedent:

• Recombinant IFN-alpha 2b is routinely used as a standard in published ELISA protocols and commercial kits for quantifying IFN-alpha in serum, plasma, and cell culture supernatants.
• Standard curves generated with recombinant IFN-alpha 2b allow for precise quantification, provided the assay is validated for this subtype.

Summary Table: Use of Recombinant IFN-alpha 2b as ELISA Standard

In conclusion, recombinant human IFN-alpha 2b is suitable and commonly used as a standard for ELISA quantification, provided you follow best practices for preparation, validation, and documentation.

Recombinant Human IFN-alpha 2b has been validated in published research for a broad range of applications, primarily in bioassays, antiviral studies, immunomodulation, and as a therapeutic agent in oncology and infectious diseases.

Key validated applications include:

• Bioactivity/Bioassays: Used extensively in cell-based assays to study its effects on immune cell activation, cytokine production, and antiviral responses.
• Antiviral Research: Demonstrated efficacy against various viruses, including hepatitis B and C, HIV-1, HPV, and SARS-CoV-2, both in vitro and in clinical studies.
• Cancer Research and Therapy: Validated for use in studies and clinical protocols for hairy cell leukemia, melanoma, follicular lymphoma, Kaposi sarcoma, chronic myelogenous leukemia, and other malignancies.
• Immunomodulation: Used to investigate modulation of immune responses, including dendritic cell activation, NK cell function, and cytokine signaling pathways (notably JAK-STAT).
• Combination Therapy Studies: Evaluated in combination with other agents (e.g., ribavirin for hepatitis, cytarabine for leukemia, Baofukang suppository for HPV infection) to assess synergistic effects and improved therapeutic outcomes.
• Prophylactic and Therapeutic Use in Infectious Disease: Studied as a prophylactic agent (e.g., intranasal administration for SARS-CoV-2 prevention) and as a treatment for persistent viral infections.
• Gynecological Applications: Used in vaginal suppository formulations for the treatment of cervical HPV infection and related gynecological conditions.

Experimental systems and sample types:

• Most validations are in human whole cell systems, but activity has also been demonstrated in murine, porcine, and bovine cell systems for cross-species antiviral studies.

Mechanistic studies:

• Frequently used to dissect JAK-STAT signaling, apoptosis, and interferon-stimulated gene expression in the context of antiviral and antitumor immunity.

Summary Table of Validated Applications

These applications are supported by both preclinical and clinical research, with broad validation in published literature for both mechanistic and therapeutic studies.

To reconstitute and prepare Recombinant Human IFN-alpha 2b protein for cell culture experiments, dissolve the lyophilized protein in sterile water or buffer at a concentration of 0.1 mg/mL (100 μg/mL), then dilute further in cell culture medium as needed for your assay.

Step-by-step protocol:

1. Preparation:

   • Briefly centrifuge the vial before opening to ensure all powder is at the bottom.
   • Warm the vial to room temperature before opening to minimize condensation.

2. Reconstitution:

   • Add sterile distilled water or sterile PBS (pH 7.4) to achieve a final concentration of 0.1 mg/mL (some protocols recommend up to 0.2 mg/mL; always check your specific product’s Certificate of Analysis).
   • If recommended, include 0.1% endotoxin-free recombinant human serum albumin (HSA) as a carrier protein to stabilize the solution, especially for long-term storage or low-concentration applications.
   • Gently swirl or tap the vial to mix; do not vortex or shake vigorously to avoid protein denaturation.

3. Dilution for Cell Culture:

   • After reconstitution, dilute the stock solution into your cell culture medium (e.g., RPMI, DMEM) containing 5–10% serum or other supplements as required for your experiment.
   • Typical working concentrations for bioassays range from picogram to nanogram per milliliter, depending on cell type and experimental design.

4. Storage:

   • Store reconstituted protein in aliquots at 4°C for up to one week or at –20°C to –80°C for longer-term storage.
   • Avoid repeated freeze-thaw cycles to preserve protein activity.
   • For extended storage, addition of carrier protein (e.g., HSA or BSA at 0.1%) is recommended.

Additional notes:

• Do not reconstitute at concentrations above 1 mg/mL to avoid solubility issues.
• Confirm protein integrity by SDS-PAGE if needed.
• Always consult the product-specific datasheet or Certificate of Analysis for buffer recommendations and activity validation.

Summary Table:

This protocol ensures optimal solubility, stability, and biological activity of recombinant IFN-alpha 2b for cell culture applications.