Recombinant Mouse IFN-γ R1

Applications and Research Value of Recombinant Mouse IFN-γR1

Recombinant mouse IFN-γR1 serves as a powerful tool for investigating interferon-gamma signaling and immune regulation in murine research models. The primary value of this recombinant protein lies in its ability to function as a high-affinity antagonist that selectively binds and neutralizes IFN-γ, allowing researchers to dissect the specific roles of this critical cytokine in various biological processes.

Key Research Applications

Immune Response Modulation

IFN-γR1 is the ligand-binding subunit essential for IFN-γ recognition and receptor internalization. By using recombinant IFN-γR1, you can block IFN-γ signaling to study how this cytokine regulates immune cell function and maturation. This is particularly valuable since IFN-γ plays central roles in Th1 immune responses, T cell differentiation, activation, and survival. Researchers can use recombinant IFN-γR1 to determine which immune outcomes are directly dependent on IFN-γ signaling versus other pathways.

Hematopoietic and Transplantation Studies

Understanding IFN-γ's role in hematopoietic stem cell (HSC) biology is critical for improving transplantation outcomes. IFN-γ reduces functional HSCs and augments Fas-mediated apoptosis in hematopoietic stem and progenitor cells. By employing recombinant IFN-γR1 to neutralize circulating IFN-γ, you can investigate mechanisms of graft rejection and develop strategies to protect transplanted cells from IFN-γ-mediated destruction.

Vaccine and Vector Development

IFN-γ suppresses antigen expression from recombinant viral vectors in a dose-dependent manner. Using recombinant IFN-γR1 to block this suppression can enhance transgene expression and improve immunogenicity of vaccine platforms, potentially leading to more effective immune responses against target antigens.

Pathogen-Specific Immunity

Since IFN-γ is essential for controlling intracellular pathogens including mycobacteria, parasites, and fungi, recombinant IFN-γR1 allows you to model scenarios where IFN-γ signaling is compromised, mimicking immunodeficiency conditions and testing therapeutic interventions.

Technical Advantages

The recombinant soluble form of IFN-γR1 can be detected in biological fluids and demonstrates species-specific binding to mouse IFN-γ. This makes it suitable for both in vitro bioassays and in vivo studies where you need to specifically antagonize IFN-γ signaling while maintaining the integrity of other cytokine pathways.

You cannot use Recombinant Mouse IFN-γ R1 as a standard for quantification or calibration in ELISA assays designed to measure IFN-γ itself; it is only appropriate as a standard for assays specifically quantifying IFN-γ R1 (the receptor), not the cytokine IFN-γ.

Key context:

• IFN-γ R1 (Interferon gamma receptor 1) is the ligand-binding chain of the IFN-γ receptor, distinct from the cytokine IFN-γ.
• ELISA kits for IFN-γ use recombinant IFN-γ as the standard to generate calibration curves for quantifying IFN-γ concentrations in samples.
• ELISA kits for IFN-γ R1 use recombinant IFN-γ R1 as the standard to quantify the receptor in biological samples.

Best practice:

• For quantifying IFN-γ in your samples, use a recombinant IFN-γ protein standard, as validated in commercial ELISA kits and protocols.
• For quantifying IFN-γ R1, use recombinant IFN-γ R1 as the standard, following protocols specific to IFN-γ R1 ELISA kits.

Summary Table:

Using the incorrect standard (e.g., IFN-γ R1 for IFN-γ quantification) will result in invalid calibration and inaccurate quantification, as the proteins are structurally and functionally distinct.

Recombinant Mouse IFN-γ R1 has been validated for several key applications in published research, primarily as a bioassay reagent, positive control, immunogen, and for use in SDS-PAGE and Western blot (WB) analyses.

Validated Applications:

• Bioassay: Recombinant Mouse IFN-γ R1 is used to assess its ability to bind IFN-γ with high affinity and act as a potent IFN-γ antagonist. This property is exploited in cell-based assays to study IFN-γ signaling and inhibition.
• Positive Control: It serves as a positive control in experiments requiring a known IFN-γ receptor protein, ensuring assay reliability and specificity.
• Immunogen: The protein is used as an immunogen for generating antibodies or for immunization protocols in animal models.
• SDS-PAGE and Western Blot: Recombinant Mouse IFN-γ R1 is validated for use in protein electrophoresis and immunoblotting, allowing researchers to confirm its presence, purity, and molecular weight.

Additional Context:

• The recombinant soluble IFN-γ R1 binds IFN-γ with high affinity and is species-specific, making it suitable for mechanistic studies of IFN-γ signaling in mouse models.
• It is commonly used in studies investigating cytokine-receptor interactions, immune modulation, and as a tool for blocking IFN-γ activity in vitro and in vivo.
• The protein's Fc fusion format can also be used in surface plasmon resonance (SPR) assays to study binding kinetics and receptor-ligand interactions.

Summary Table

These applications are supported by published research and product documentation, confirming the protein's utility in immunological, biochemical, and cell biology studies.

To reconstitute and prepare Recombinant Mouse IFN-γ R1 protein for cell culture experiments, dissolve the lyophilized protein in sterile phosphate-buffered saline (PBS) at a concentration of 100 μg/mL. For optimal stability and activity in cell culture, add a carrier protein such as 0.1% Bovine Serum Albumin (BSA) to the solution.

Step-by-step protocol:

• Before opening the vial: Briefly centrifuge the vial (20–30 seconds in a microcentrifuge) to collect all lyophilized powder at the bottom.
• Reconstitution: Add sterile PBS to achieve the desired concentration (typically 100 μg/mL for IFN-γ R1). Gently pipette to dissolve the protein, washing down the sides of the vial to ensure complete recovery.
• Carrier protein: Include 0.1% BSA in the PBS to enhance protein stability and prevent adsorption to tube walls, especially for long-term storage or dilute solutions.
• Mixing: Do not vortex or mix vigorously. Gently invert or pipette to mix.
• Incubation: If solubility issues arise, allow the solution to incubate at 4 °C for several hours or overnight.
• Aliquoting and storage: After reconstitution, aliquot the solution to avoid repeated freeze-thaw cycles. Store aliquots at −20 °C to −70 °C for long-term use, or at 2–8 °C for up to 60 days.
• Quality check: Confirm protein recovery by SDS-PAGE if necessary.

Additional notes:

• For cell culture experiments, dilute the reconstituted stock into your complete cell culture medium immediately before use.
• Avoid using lysis buffers containing surfactants or organic solvents for preparation, as these may denature the protein.
• If using a carrier-free formulation, ensure that the absence of BSA will not affect your downstream applications.

Summary of key points:

• Buffer: Sterile PBS
• Concentration: 100 μg/mL (adjust as needed)
• Carrier protein: 0.1% BSA recommended
• Storage: Aliquot and freeze at −20 °C to −70 °C; short-term at 2–8 °C
• Handling: Gentle mixing, avoid vortexing

This protocol ensures the recombinant Mouse IFN-γ R1 protein is properly reconstituted and stabilized for reliable use in cell culture assays.