Anti-Mouse CD120a (TNFR1) [Clone 55R-593] — Purified in vivo PLATINUM™ Functional Grade

Common In Vivo Applications of Clone 55R-593 in Mice

Clone 55R-593 is a monoclonal antibody that specifically targets the mouse CD120a antigen, also known as TNFR1 (Tumor Necrosis Factor Receptor type I). This clone is widely used in vivo in mouse models for the following purposes:

• Specific Blockade of TNFR1 Signaling: Clone 55R-593 is primarily used to block TNFR1 signaling in vivo, allowing researchers to investigate the specific role of this receptor in inflammation, immunity, and various disease models. By blocking TNFR1, scientists can dissect the contributions of this pathway independent of other TNF receptor pathways.
• Inflammation and Disease Modeling: The antibody is used to study the effects of TNF signaling in models of inflammatory diseases (e.g., arthritis, colitis), autoimmune disorders, and other conditions where TNFα is a key mediator. This helps in understanding the mechanisms of disease and evaluating potential therapeutic interventions targeting this receptor.
• Apoptosis and Cell Survival Studies: Since TNFR1 activation can trigger apoptosis through its death domain, clone 55R-593 is employed to study cell death mechanisms in vivo, especially in contexts where TNFα-induced cytotoxicity is relevant.
• NF-κB Activation and Gene Expression: The antibody can be used to modulate the downstream effects of TNFR1 activation, including NF-κB signaling and the expression of proinflammatory genes, thus elucidating the role of this pathway in immune responses and tissue damage.

Mechanism and Specificity

Clone 55R-593 recognizes mouse CD120a (TNFR1), is of Armenian Hamster IgG isotype, and binds the extracellular domain of the receptor. It blocks ligand-induced receptor signaling when administered in vivo, effectively neutralizing the biological activity of TNFα and TNFβ (LTα) at the receptor level. This specificity is critical for experiments designed to isolate the effects of TNFR1 from those of other TNF receptors.

Technical Considerations

• Functional Grade: This antibody is available in highly purified, low-endotoxin, and functional grade formats (e.g., In Vivo GOLD™, In Vivo PLATINUM™), making it suitable for sensitive in vivo applications without eliciting non-specific immune responses.
• No Cross-Reactivity: Clone 55R-593 does not cross-react with TNFR2, ensuring that observed effects are specifically due to TNFR1 blockade.
• Applications Beyond Blocking: While its primary in vivo application is blocking, the antibody is also validated for immunoprecipitation and Western blotting in research contexts.

Summary Table

Key Point

Clone 55R-593 is a critical tool for in vivo studies aiming to specifically block TNFR1 signaling in mice, enabling precise investigation of TNFα biology in health and disease.

The 55R-593 antibody, targeting mouse CD120a (TNFR1), is commonly used in various applications such as immunoprecipitation, receptor blocking, and as a TNF-α agonist. In the literature, it is often used in conjunction with other antibodies and proteins to study signaling pathways, apoptosis, and inflammation. Some of these commonly used antibodies and proteins include:

• Signaling Proteins:

  • Anti-phospho-NF-κB and Anti-phospho-ERK: These are used for signaling pathway analysis to study the downstream effects of TNF-α signaling through TNFR1.

• Apoptosis Markers:

  • Anti-cleaved caspase-3 and Annexin V: These are used in cell death assays to examine apoptosis, which is a potential outcome of TNF-α signaling.

• Inflammatory Markers:

  • Anti-ICAM-1 and Anti-VCAM-1: These are used in inflammation studies, as TNF-α is a pro-inflammatory cytokine that can induce the expression of adhesion molecules like ICAM-1 and VCAM-1.

Additionally, the 55R-593 antibody may be used alongside other antibodies targeting related proteins in the TNF pathway or in studies of inflammation and immune response. However, specific literature references to these combinations are not detailed in the search results.

Clone 55R-593, an Armenian Hamster IgG monoclonal antibody targeting mouse CD120a (TNFR1), has been used in various scientific studies. Here are some key findings from citations involving this clone:

• Agonist Activity: Unlike other anti-p55 antibodies, clone 55R-593 functions as an agonist, triggering cell lysis, nitric oxide induction, and activating NF-κB.
• Application in Research: Clone 55R-593 has been used to investigate the role of TNF receptors in certain biological processes, such as the effects of IL-33.
• Receptor Blocking and Agonist Use: The antibody is useful for immunoprecipitation, in vitro and in vivo receptor blocking, and as an agonist of TNF-α.
• Specificity and Expression: Clone 55R-593 specifically recognizes mouse CD120α, which is constitutively expressed in most tissues at low levels.

These findings highlight the versatility and specificity of clone 55R-593 in studying TNF signaling and related biological processes.

Dosing regimens of clone 55R-593 (anti-mouse CD120a/TNFR1) in mouse models are highly variable and are influenced by several key factors: the specific experimental goal (receptor blockade vs. agonist activity), the mouse strain or disease model, and sometimes the application route. There is no universally standardized dosing protocol for this antibody across all mouse models.

Key contextual points:

• Experimental Purpose: For receptor blockade, researchers often titrate the dose based on desired level of TNFR1 inhibition, while for agonist experiments (stimulating activity through TNFR1), higher or more frequent dosing may be needed.
• Mouse Strain & Disease Context: Strain sensitivities (e.g., BALB/c vs. C57BL/6) and disease contexts (inflammation, autoimmunity, tumor models) may require adjustment of doses to optimize efficacy and minimize off-target effects.
• Reported Practice: Most suppliers and technical data sheets, including Leinco and BioLegend, emphasize empirical optimization of dosing: protocols are often adapted or titrated to the specific biology of the model and experimental readout rather than following a single published standard.
• Application Examples: The antibody is used both for functional blockade (in vivo administration), and for immune assays (in vitro and in vivo), but precise μg/mouse or mg/kg numbers are not consistently detailed in the main supplier or product literature—unlike more commonly used clones such as those targeting PD-1 or CTLA-4.

Summary Table: Factors Influencing Dosing Regimen for Clone 55R-593

Notable Gaps:
Unlike widely cited in vivo mAb protocols for checkpoint blockade (PD-1, PD-L1, CTLA-4) which recommend 100-500 μg per mouse 2-3 times a week, published or manufacturer protocols for clone 55R-593 do not specify a widely accepted standard dose or schedule. Investigators are advised to begin with dose-ranging pilot studies tailored to their specific experimental question and mouse model, monitoring both efficacy (receptor engagement/blockade) and potential toxicity.

Conclusion:
Dosing of clone 55R-593 is not standardized and should be empirically determined for each experimental setting, with protocol adaptations based on the biology of the system under investigation and the goals of the experiment.