Recombinant Rat IFNγ

Recombinant Rat IFNγ (Interferon gamma) is a valuable tool for research applications due to its well-characterized biological functions and its critical role in immune regulation. Here are several reasons why you should consider using Recombinant Rat IFNγ in your research:

1. Immune Modulation and Activation

• IFNγ is a key cytokine produced by activated T cells and natural killer (NK) cells.
• It modulates macrophage effector functions, enhances antigen presentation by upregulating MHC class I and II molecules, and promotes the activation of dendritic cells and mononuclear phagocytes.
• IFNγ also stimulates the production of IL-12 by dendritic cells, further amplifying immune responses.

2. Antiviral and Antiproliferative Effects

• IFNγ exhibits strong antiviral activity and can inhibit the proliferation of various cell types, including tumor cells.
• It is involved in the induction of class I and II MHC molecules, which are essential for immune surveillance and the elimination of infected or malignant cells.

3. Regulation of Immune Cell Differentiation

• IFNγ plays a crucial role in the development and activation of Th1 cells, which are important for cell-mediated immunity.
• It also influences isotype switching and potentiates the secretion of immunoglobulins by B cells, contributing to humoral immunity.

4. Standardization and Reliability

• Recombinant Rat IFNγ is produced under controlled conditions, ensuring high purity and consistency.
• This standardization is essential for reproducible results in functional studies, binding assays, and therapeutic development programs.

5. Versatile Applications

• Recombinant Rat IFNγ can be used in a wide range of applications, including:
  • Bioassays to study immune cell activation and function.
  • Functional studies to investigate the effects of IFNγ on various cell types.
  • Binding assays to explore receptor interactions.
  • Therapeutic development to evaluate the potential of IFNγ in treating diseases.

6. Species-Specificity

• The biological activity of IFNγ is highly species-specific. Using Recombinant Rat IFNγ ensures that your experiments are relevant to rat models, which is particularly important for translational research.

7. Research in Disease Models

• IFNγ is involved in the pathogenesis of various diseases, including autoimmune disorders, infections, and cancer. Using Recombinant Rat IFNγ allows you to study these processes in rat models, providing insights into disease mechanisms and potential therapeutic targets.

8. High-Quality Reagents

• Many suppliers offer Recombinant Rat IFNγ with high purity and activity, making it a reliable reagent for advanced research applications in molecular biology and biotechnology.

In summary, Recombinant Rat IFNγ is a powerful and versatile tool for studying immune responses, disease mechanisms, and therapeutic interventions in rat models. Its well-defined biological functions and high-quality production make it an essential reagent for a wide range of research applications.

Yes, recombinant rat IFNγ can be used as a standard for quantification or calibration in ELISA assays, provided it is properly validated for this purpose. Many commercial ELISA kits for rat IFNγ use recombinant protein as the standard and demonstrate that the assay quantifies both recombinant and natural forms equivalently.

Key considerations and supporting details:

• Assay Compatibility: Most rat IFNγ ELISA kits are designed to detect both natural and recombinant forms of the cytokine, and their standard curves are typically generated using recombinant rat IFNγ. This ensures that the quantification is accurate for both sample types.

• Parallelism: Validation data from reputable kits show that dose-response curves for recombinant and natural rat IFNγ are parallel, indicating that the recombinant protein is suitable as a standard for quantification.

• Standard Preparation: When using recombinant rat IFNγ as a standard, it is critical to:

  • Reconstitute and dilute the protein according to the manufacturer’s or protocol’s instructions.
  • Use the same buffer or diluent as recommended for the assay to avoid matrix effects.
  • Prepare a standard curve covering the assay’s dynamic range (e.g., 8–500 pg/mL or as specified by your kit).

• No International Reference Standard: There is currently no international reference standard for rat IFNγ, so quantification is relative to the recombinant standard used in your assay. This is standard practice in cytokine ELISAs.

• Documentation: Some suppliers specifically market recombinant rat IFNγ as an ELISA standard, confirming its intended use for calibration in sandwich ELISA protocols.

• Validation: If you are using a custom or non-kit recombinant IFNγ, ensure it is full-length, correctly folded, and biologically active. Ideally, compare its standard curve to that of a validated kit standard to confirm equivalence.

Summary Table: Recombinant Rat IFNγ as ELISA Standard

In conclusion, recombinant rat IFNγ is appropriate for use as a standard in ELISA quantification, provided it is validated and prepared according to best practices.

Recombinant Rat IFNγ has been validated in published research primarily for bioassays in cell culture and in vivo studies to investigate its immunomodulatory, antiviral, and pro-inflammatory activities. The most common applications include:

• Bioassays (in vitro):

  • Activation of macrophages and monocytes to study immune responses.
  • Induction of MHC class I and II expression on various cell types.
  • Investigation of cytokine-induced apoptosis and necrosis in pancreatic beta cells.
  • Analysis of NOX-1 expression and signaling pathways in beta cells.
  • Modeling inflammatory and neuropathic pain mechanisms in astrocytes.
  • Studying wound healing and immunomodulation in tissue culture models.
  • Assessing anti-parasitic and antiviral responses in infected cell cultures.

• In vivo applications:

  • Intratumoral infusion to study immune cell infiltration and tumor microenvironment modulation in rat glioma models.
  • Intranasal administration to evaluate antiviral protection against pathogens such as SARS-CoV-2 in animal models (primarily mouse, but mechanistically relevant to rat studies).
  • Investigation of wound healing and tissue regeneration in rat models.

• Other validated uses:

  • As a standard in immunoassays (e.g., ELISA) for quantifying IFNγ activity or concentration.
  • Functional assays to assess cytokine bioactivity in various immune cell types.

Key research areas and protocols:

• Immunology: Dissecting Th1 cell development, macrophage activation, and cytokine signaling.
• Neuroscience: Modeling neuroinflammation and neuropathic pain.
• Diabetes research: Studying beta-cell apoptosis and immune-mediated islet damage.
• Cancer biology: Evaluating tumor immune responses and cytokine-induced tumor invasion.
• Infectious disease: Assessing antiviral and anti-parasitic mechanisms.

Summary Table:

References to published research confirm these applications, with studies using recombinant rat IFNγ to probe immune mechanisms, disease models, and therapeutic interventions in both cell culture and animal models.

To reconstitute and prepare Recombinant Rat IFNγ protein for cell culture experiments:

• Centrifuge the vial briefly before opening to ensure all lyophilized material is at the bottom.
• Add sterile water to achieve a concentration of 0.1 mg/mL (100 µg/mL) unless your protocol specifies otherwise. Some protocols also allow reconstitution in sterile PBS with at least 0.1% BSA or HSA to stabilize the protein and prevent adsorption to plastic.
• Gently pipette the solution up and down to dissolve the protein. Do not vortex, as this can denature the protein.
• Allow several minutes for complete reconstitution at room temperature.
• If precipitate is observed, centrifuge the solution and use only the clear, soluble fraction for experiments.
• For long-term storage, aliquot the reconstituted protein in a buffer containing 0.1% BSA and store at -80°C to avoid repeated freeze-thaw cycles.
• For short-term use, store at 4°C for up to one week.

Summary of best practices:

Additional notes:

• Always use sterile technique to avoid contamination.
• If using for sensitive cell types, ensure the final buffer is compatible with your cell culture system.
• Avoid repeated freeze-thaw cycles by aliquoting.

These steps will help maintain the biological activity and stability of recombinant rat IFNγ for reliable cell culture experiments.