Recombinant Mouse IFN-γ R1

Applications and Research Value of Recombinant Mouse IFNγR1

Recombinant mouse IFNγR1 serves as a powerful tool for investigating interferongamma signaling and immune regulation in murine research models. The primary value of this recombinant protein lies in its ability to function as a highaffinity antagonist that selectively binds and neutralizes IFNγ, allowing researchers to dissect the specific roles of this critical cytokine in various biological processes.

Key Research Applications

Immune Response Modulation

IFNγR1 is the ligandbinding subunit essential for IFNγ recognition and receptor internalization. By using recombinant IFNγR1, you can block IFNγ signaling to study how this cytokine regulates immune cell function and maturation. This is particularly valuable since IFNγ plays central roles in Th1 immune responses, T cell differentiation, activation, and survival. Researchers can use recombinant IFNγR1 to determine which immune outcomes are directly dependent on IFNγ signaling versus other pathways.

Hematopoietic and Transplantation Studies

Understanding IFNγ's role in hematopoietic stem cell (HSC) biology is critical for improving transplantation outcomes. IFNγ reduces functional HSCs and augments Fasmediated apoptosis in hematopoietic stem and progenitor cells. By employing recombinant IFNγR1 to neutralize circulating IFNγ, you can investigate mechanisms of graft rejection and develop strategies to protect transplanted cells from IFNγmediated destruction.

Vaccine and Vector Development

IFNγ suppresses antigen expression from recombinant viral vectors in a dosedependent manner. Using recombinant IFNγR1 to block this suppression can enhance transgene expression and improve immunogenicity of vaccine platforms, potentially leading to more effective immune responses against target antigens.

PathogenSpecific Immunity

Since IFNγ is essential for controlling intracellular pathogens including mycobacteria, parasites, and fungi, recombinant IFNγR1 allows you to model scenarios where IFNγ signaling is compromised, mimicking immunodeficiency conditions and testing therapeutic interventions.

Technical Advantages

The recombinant soluble form of IFNγR1 can be detected in biological fluids and demonstrates speciesspecific binding to mouse IFNγ. This makes it suitable for both in vitro bioassays and in vivo studies where you need to specifically antagonize IFNγ signaling while maintaining the integrity of other cytokine pathways.

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

Key context:

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

Best practice:

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

Summary Table:

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

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

Validated Applications:

• Bioassay: Recombinant Mouse IFNγ R1 is used to assess its ability to bind IFNγ with high affinity and act as a potent IFNγ antagonist. This property is exploited in cellbased assays to study IFNγ signaling and inhibition.
• Positive Control: It serves as a positive control in experiments requiring a known IFNγ 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.
• SDSPAGE and Western Blot: Recombinant Mouse IFNγ 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 IFNγ R1 binds IFNγ with high affinity and is speciesspecific, making it suitable for mechanistic studies of IFNγ signaling in mouse models.
• It is commonly used in studies investigating cytokinereceptor interactions, immune modulation, and as a tool for blocking IFNγ 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 receptorligand 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 IFNγ R1 protein for cell culture experiments, dissolve the lyophilized protein in sterile phosphatebuffered 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.

Stepbystep 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 IFNγ 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 longterm 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 freezethaw cycles. Store aliquots at −20 °C to −70 °C for longterm use, or at 2–8 °C for up to 60 days.
• Quality check: Confirm protein recovery by SDSPAGE 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 carrierfree 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; shortterm at 2–8 °C
• Handling: Gentle mixing, avoid vortexing

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