Anti-Mouse IFNß – Purified in vivo PLATINUM™ Functional Grade

In Vivo Applications of Clone HDß-4A7 in Mice

Clone HDß-4A7 is a monoclonal antibody specifically targeting murine interferon-beta (IFN-β), a key cytokine in the innate immune response. In vivo, this antibody is primarily used to neutralize or block IFN-β signaling in mice, enabling researchers to investigate the functional consequences of IFN-β deficiency during infection and immune challenge.

Specific In Vivo Uses

• Modeling IFN-β Deficiency: Administration of HDß-4A7 in mice effectively mimics the phenotype of IFN-β knockout mice, allowing study of the specific role of IFN-β in immune responses without the need for genetic modification.
• Viral Pathogenesis Studies: A prominent application is in the study of West Nile Virus (WNV) infection. Blocking IFN-β with HDß-4A7 increases the lethality of WNV in mice, mirroring the increased susceptibility seen in IFN-β-deficient mice, thus highlighting the protective role of endogenous IFN-β against viral infection.
• Investigating Innate Immunity: By transiently neutralizing IFN-β, researchers can dissect its contributions to early antiviral defenses, cytokine cascades, and the regulation of other immune cells and pathways in vivo.

Technical and Experimental Design

• Dosing and Specificity: HDß-4A7 binds both recombinant and natural murine IFN-β and does not cross-react with other interferons such as IFN-γ or multiple IFN-α species, ensuring specificity in experimental outcomes.
• Functional Readouts: The consequences of IFN-β blockade can be assessed via survival curves, viral load measurements, immune cell profiling, and cytokine analyses in challenged mice.
• Complementary Tools: HDß-4A7 is sometimes used alongside other antibodies that block the IFN-α/β receptor (e.g., MAR1-5A3), allowing researchers to distinguish between the effects of IFN-α and IFN-β or to achieve complete type I interferon signaling blockade.

Summary Table: Key Applications

Conclusions

Clone HDß-4A7 is a crucial tool for in vivo research in mice, primarily used to neutralize IFN-β and study its unique role in antiviral immunity and innate immune regulation. Its applications are especially valuable in viral infection models, where IFN-β's protective effects are a major focus, and in studies aiming to dissect the specific contributions of IFN-β apart from other interferons.

Commonly used antibodies or proteins with HDß-4A7 (an antibody specific for murine interferon-beta, IFN-β) in the literature include:

• Anti-IFNAR1 mAb (MAR1-5A3): Frequently used as a comparison or in combination, since it blocks the type I interferon receptor, inhibiting signaling from IFN-β and all type I interferons.
• MHC Class I (H2-Kb) detection antibodies: Used to assess IFN-β–induced upregulation of MHC-I, providing a readout for IFN-β activity and HDß-4A7 blocking efficiency.
• Stat1 phosphorylation detection (e.g., anti-pStat1 antibodies): Utilized to monitor IFN-β–mediated signaling, where HDß-4A7 inhibition can show its specificity by reducing Stat1 activation in response to IFN-β.

Additional proteins and controls often included in these experiments:

• Murine IFN-γ and various IFN-α subtypes: Used as controls to confirm the specificity of HDß-4A7 for IFN-β—it does not bind IFN-γ or several IFN-α species.
• Recombinant and natural forms of IFN-β: Used in binding and functional assays to demonstrate specificity and neutralizing capability of HDß-4A7.
• VSV (Vesicular Stomatitis Virus): Frequently used as a viral challenge in functional assays to test the antiviral effect mediated by IFN-β and its inhibition by HDß-4A7.

In summary, anti-IFNAR1, MHC-I (H2-Kb), anti-pStat1, and various type I interferons are commonly used in concert with HDß-4A7. These combinations are employed to dissect the specificity, signaling pathways, and biological effects mediated by IFN-β in murine models.

Key Findings from Clone HDß-4A7 Antibody Research

Clone HDß-4A7 is a mouse monoclonal IgG2a antibody developed to specifically target and bind murine interferon-beta (IFN-β), a key type I interferon involved in innate immunity and antiviral responses.

In Vitro Functions

• Selective Binding: HDß-4A7 recognizes an epitope on mouse IFN-β and does not bind to mouse IFN-γ or several different forms of IFN-α (α-1, 4, 5, or 13), demonstrating high specificity for IFN-β.
• Functional Neutralization: The antibody inhibits IFN-β-induced STAT1 phosphorylation in cell lines, with a potency similar to that of an anti-IFNAR1 antibody (MAR1-5A3).
• Downstream Effects: HDß-4A7 blocks the IFN-β-induced upregulation of major histocompatibility complex class I (MHC-I, H2-Kb) expression in fibrosarcoma and L929 cell lines.
• Antiviral Activity: It significantly neutralizes the antiviral activity of IFN-β in vitro, as shown in experiments with L929 cells infected with vesicular stomatitis virus (VSV).

In Vivo Applications

• Biological Activity: When administered in vivo, HDß-4A7 circulates with a half-life of approximately 14 days in mice.
• Phenotypic Mimicry: The antibody effectively blocks endogenous IFN-β signaling, leading to increased lethality in mice infected with West Nile virus (WNV), recapitulating the phenotype observed in mice genetically deficient for IFN-β (IFNβ-/-).
• Manufacturing Standards: The antibody is produced in an animal-free facility using in vitro cell culture and is purified to ensure low levels of endotoxins, protein A/G, and aggregates, making it suitable for sensitive in vivo studies.

Research Applications

• Modeling IFNβ Deficiency: HDß-4A7 is used to transiently model IFN-β deficiency in vivo, allowing for the study of IFN-β's role in infection and immunity without genetic knockout.
• Experimental Flexibility: Researchers are advised to determine optimal working concentrations for their specific assays, as antibody performance may vary by application.

Summary Table: Key Properties and Effects of HDß-4A7

Conclusion

Clone HDß-4A7 is a well-characterized, highly specific tool for studying the biological roles of mouse IFN-β, both in vitro and in vivo. Its ability to functionally neutralize IFN-β allows researchers to investigate the consequences of transient IFN-β blockade, particularly in models of viral infection and immune regulation.

There are currently no published dosing regimens for the antibody clone HDß-4A7 in mouse models. Standard references specializing in in vivo antibody dosing for mice do not list recommendations or examples for this specific clone. As such, there is no authoritative guidance on how dosing of HDß-4A7 varies across different mouse strains, disease models, or experimental applications.

Dosing practices for other antibody clones (e.g., checkpoint inhibitors like anti-PD-1, anti-PD-L1, anti-CTLA-4) commonly depend on:

• Mouse strain and age
• Disease context (e.g., tumor model, infection, autoimmune disease)
• Experimental goal (e.g., depletion, blockade, activation)
• Administration route (usually intraperitoneal injection)
• Dose range (often 0.1–0.5 mg per mouse for functional antibodies)
• Frequency (every 3–7 days, or tailored to model kinetics)

In the absence of data for HDß-4A7, researchers typically follow the approach of starting with dose ranges and regimens established for antibodies with similar targets and purposes, then optimize for their specific experimental scenario. For new or uncharacterized antibodies, pilot titration studies and literature review of related clones are recommended.

If you require a dosing regimen for HDß-4A7:

• Consider contacting the supplier for unpublished recommendations.
• Review literature for the dosing of antibodies against similar targets.
• Conduct preliminary dosing and titration studies tailored to your experimental mouse model.

No source currently provides published protocols or variable guidelines for clone HDß-4A7 in any mouse model.