Recombinant Human IFN-γ R1

Recombinant Human IFNγ R1 is primarily used in research as a tool to study and modulate interferongamma (IFNγ) signaling, serving as a highaffinity, speciesspecific ligandbinding subunit that can act as a decoy or antagonist for IFNγ in various experimental systems.

Key reasons to use recombinant human IFNγ R1 in research applications include:

• Functional Antagonist: Soluble recombinant IFNγ R1 binds IFNγ with high affinity, effectively sequestering the cytokine and preventing it from interacting with cellsurface receptors. This allows researchers to specifically inhibit IFNγmediated signaling in vitro or in vivo, enabling the study of IFNγ’s biological roles by observing the effects of its blockade.
• Mechanistic Studies: By blocking IFNγ signaling, you can dissect the contribution of this pathway to immune responses, inflammation, cell proliferation, apoptosis, and host defense mechanisms. This is particularly useful in models of infection, autoimmunity, cancer, and fibrosis, where IFNγ plays a central regulatory role.
• Receptor Biology: Recombinant IFNγ R1 can be used to characterize ligandreceptor interactions, determine binding kinetics, and map functional domains involved in cytokine recognition and receptor activation.
• Assay Development: It serves as a standard or competitor in bioassays designed to measure IFNγ activity, screen for IFNγ inhibitors, or validate antibody specificity in blocking IFNγ signaling.

Technical context:

• IFNγ R1 is the essential ligandbinding subunit of the IFNγ receptor complex; it is necessary and sufficient for IFNγ binding and receptor internalization, while IFNγ R2 is required for downstream signaling but does not bind IFNγ independently.
• Recombinant soluble IFNγ R1 is typically produced as the extracellular domain, which retains highaffinity binding to IFNγ and is detected in biological fluids as a natural antagonist.
• Human and mouse IFNγ R1 are only partially homologous and bind IFNγ in a speciesspecific manner, so it is important to match the recombinant protein to the species of IFNγ used in your experiments.

Common applications:

• Blocking IFNγ activity in cell culture or animal models to study immune regulation, cytokine networks, or disease pathogenesis.
• Investigating the therapeutic potential of IFNγ antagonism in models of autoimmunity, chronic inflammation, or cytokine storm.
• Validating the specificity of IFNγ neutralizing antibodies or smallmolecule inhibitors.

In summary, recombinant human IFNγ R1 is a valuable reagent for selectively inhibiting IFNγ signaling, dissecting cytokine function, and developing assays or therapeutics targeting the IFNγ pathway.

Recombinant Human IFNγ R1 (Interferon Gamma Receptor 1) should not be used as a standard for quantification or calibration in ELISA assays designed to measure IFNγ itself. ELISA standards must match the analyte being quantified; IFNγ R1 is a receptor protein, not the cytokine IFNγ.

Supporting details:

• ELISA quantification standards are typically recombinant or purified forms of the analyte of interest—in this case, human IFNγ—not its receptor. These standards are calibrated against international reference preparations (e.g., NIAID or NIBSC standards) to ensure accurate quantification of IFNγ concentrations in biological samples.
• Recombinant Human IFNγ R1 is used as a standard only in ELISA kits specifically designed to measure IFNγ R1 levels, such as in studies of receptor expression, shedding, or disease biomarker research. It is not interchangeable with IFNγ for cytokine quantification.
• Using IFNγ R1 as a standard in an IFNγ ELISA would result in inaccurate calibration, as the antibodies in IFNγ ELISA kits are specific for the cytokine, not its receptor.

Best practice:

• For IFNγ quantification, use a recombinant human IFNγ protein standard, calibrated against an international reference, as provided or recommended by your ELISA kit manufacturer.
• For IFNγ R1 quantification, use recombinant IFNγ R1 as a standard in receptorspecific ELISA kits.

Summary Table:

In conclusion:
Use recombinant human IFNγ R1 only as a standard for assays measuring IFNγ R1, not for IFNγ quantification ELISAs. For IFNγ ELISA calibration, use recombinant IFNγ protein standards.

Recombinant Human IFNγ R1 has been validated primarily for use in bioassays as a soluble receptor and antagonist of IFNγ signaling in published research. Its main applications include:

• Bioassays: Used to assess its ability to bind IFNγ with high affinity and antagonize IFNγ activity in cellbased systems.
• Binding studies: Validated for measuring direct interaction with IFNγ, confirming its function as a decoy receptor.
• Functional inhibition: Employed to block IFNγmediated cellular responses, such as cytokine signaling, gene expression, and immune activation, in various cell types.

Supporting details:

• The recombinant soluble IFNγ R1 has been shown to bind IFNγ with high affinity and act as a potent antagonist, inhibiting IFNγdriven biological effects in vitro.
• Published studies have used it to dissect IFNγ signaling pathways, evaluate the specificity of IFNγ responses, and explore therapeutic strategies for diseases involving IFNγ dysregulation.
• It is also referenced in research on heparan sulfate mimicry, where it is used to demonstrate inhibition of IFNγ activity by synthetic glycoconjugates.

Additional relevant applications (based on broader research use):

• ELISA standard: Recombinant IFNγ R1 can be used as a standard or control in immunoassays for quantifying IFNγ or its receptor, though direct published validation for this specific recombinant protein is less frequently cited.
• Cell culture experiments: Used to modulate IFNγ signaling in cultured cells to study immune responses or disease models.

No evidence was found in the provided results for use in applications such as immunohistochemistry, flow cytometry, or in vivo animal studies for this specific recombinant protein. The most robust validation remains in bioassays and binding/inhibition studies.

To reconstitute and prepare Recombinant Human IFNγ R1 protein for cell culture experiments, dissolve the lyophilized protein in sterile phosphatebuffered saline (PBS) containing at least 0.1% carrier protein, such as human or bovine serum albumin (HSA or BSA), to a final concentration of 500 μg/mL. This approach minimizes protein loss due to adsorption and stabilizes the protein for downstream applications.

Stepbystep protocol:

• Before opening the vial: Briefly centrifuge the vial to collect all lyophilized material at the bottom.
• Reconstitution: Add sterile PBS (pH 7.2–7.4) containing at least 0.1% carrier protein (HSA or BSA) to achieve the desired concentration (commonly 500 μg/mL for stock solutions).
• Mixing: Gently swirl or tap the vial to dissolve the protein. Avoid vigorous pipetting or vortexing, which may denature the protein.
• Aliquoting: Divide the reconstituted protein into singleuse aliquots to avoid repeated freezethaw cycles, which can reduce activity and stability.
• Storage: Store aliquots at −20°C or −80°C. Avoid repeated freezethaw cycles.

Additional notes:

• If carrierfree protein is required, reconstitute in sterile PBS alone, but use immediately and avoid adsorption to plastic surfaces.
• For cell culture experiments, further dilute the stock solution in cell culture medium or buffer containing carrier protein to the working concentration required for your assay.
• Confirm protein concentration and integrity by SDSPAGE or other suitable analytical methods if necessary.

Summary of key points:

• Use sterile PBS + 0.1% HSA/BSA for reconstitution.
• Target 500 μg/mL for stock solution.
• Aliquot and freeze to avoid repeated freezethaw.
• Gently mix to dissolve, do not vortex.

This protocol ensures optimal stability and bioactivity of recombinant human IFNγ R1 for cell culture applications.