Anti-Mouse CD120a (TNFR1) [Clone 55R-170] — Purified in vivo PLATINUM™ Functional Grade

The monoclonal antibody clone 55R-170 is primarily used in in vivo mouse studies for several applications:

1. Neutralization of Mouse TNF Receptor I (TNFRI) Bioactivity: This antibody is used to block the bioactivity of TNFRI, which is crucial for studying the role of TNFRI-mediated signaling in models of inflammation, infection, and TNF-related pathology.

2. Protection from Lethal Endotoxin Shock: Clone 55R-170 has been shown to protect mice from lethal endotoxin shock by inhibiting TNF-alpha effects, which indicates its utility in studying sepsis and other inflammatory conditions.

3. Blocking Development of Protective Responses: It can block the development of protective responses against infections like Listeria monocytogenes, providing insights into immune mechanisms.

4. Study of Neurodegenerative Diseases: TNFRI, targeted by 55R-170, is involved in neurodegenerative diseases such as Parkinson's and Alzheimer's, allowing researchers to explore the role of inflammation in these diseases.

5. Blocking Skin Necrosis: Both p55 and p75 TNF receptor-specific mAbs, including 55R-170, can individually block the development of skin necrosis in mice treated with murine TNF-alpha.

Commonly used antibodies and proteins paired with 55R-170 (anti-mouse CD120a/TNFR1) in the literature include:

• 55R-286: Frequently paired for ELISA assays, where 55R-170 serves as the detection antibody to complement 55R-286 as the capture antibody.
• 55R-176: Used in functional studies alongside 55R-170 to compare TNFR1 activity and signaling.
• Anti-p75: Often included for comparative analyses to distinguish TNFR1 (p55) from TNFR2 (p75) signaling pathways and biological functions.
• Biotinylated detection systems: Such as streptavidin-phycoerythrin (SAv-PE) and biotin anti-mouse antibodies used in flow cytometry alongside 55R-170.
• Isotype controls: Armenian hamster IgG is commonly used as an isotype control for specificity in immunological assays with 55R-170.
• Detection reagents for flow cytometry: Anti-Armenian hamster IgG secondary antibodies are used with 55R-170 in multicolor flow cytometry setups.
• Proteins studied in TNFR1 signaling: While not direct antibody pairs, proteins such as TRAF2, TRADD, and BAG4 (SODD) are frequently analyzed in studies using 55R-170 to interrogate TNFR1-mediated effects.

These combinations enable detailed analysis of TNFR1 signaling, comparative assessment between TNFR1 and TNFR2, and precise quantification in ELISA and flow cytometry protocols. For specific experimental setups or additional pairing strategies, individual publications and manufacturer datasheets should be consulted.

Key Findings from Clone 55R-170 (Anti-Mouse CD120a/TNFR1) in Scientific Literature

Clone 55R-170 is a well-characterized monoclonal antibody targeting the mouse CD120a antigen, also known as TNFR1 (Tumor Necrosis Factor Receptor 1), and has played a significant role in elucidating the biological functions of TNF signaling in mice.

Functional Characterization

• Specific Antagonism: 55R-170 is a potent antagonist of mouse TNFR1, selectively blocking TNFR1-mediated signaling both in vitro and in vivo. This specificity has been leveraged to dissect the distinct roles of TNFR1 and TNFR2 (CD120b) in inflammation, infection, and cytokine storm.
• In Vitro Inhibition: At a concentration of 25 µg/mL, 55R-170 inhibits by 50% the biological effects of 1 ng/mL mouse TNF-α in cytotoxicity assays using L929 cells, demonstrating its efficacy in neutralizing TNF bioactivity.
• Ligand Binding Blockade: Functional studies show that 55R-170, along with clone 55R-176, blocks TNF binding to TNFR1 and acts as a TNF antagonist, in contrast to another clone (55R-593) which exhibits agonist activity.
• In Vivo Applications: When administered in vivo, anti-p55 (TNFR1) antibodies like 55R-170 protect mice from lethal endotoxin shock and block the development of a protective immune response against Listeria monocytogenes infection, highlighting the critical role of TNFR1 in these processes.
• Distinct from TNFR2 Effects: Blocking TNFR1 (but not TNFR2) with 55R-170 influences outcomes in models of endotoxin shock and infection, while both receptors contribute to the development of TNF-induced skin necrosis.

Research Applications

• Mechanistic Insights: The use of 55R-170 has provided mechanistic insights into how TNFR1 signaling contributes to disease pathogenesis, informing therapeutic hypotheses for conditions involving dysregulated TNF signaling.
• Tool for Immunotherapy Research: 55R-170 has been used in combination with other immunotherapies (e.g., anti-PD-1) to explore synergistic effects in cancer models, suggesting potential for combination strategies in modulating anti-tumor immunity.
• Assay Utility: Beyond in vivo studies, 55R-170 is validated for ELISA, flow cytometry, and bioassays, and is available in various functional formats (e.g., biotinylated for detection).

Technical Specifications

• High Purity: The antibody is produced at over 90% purity and with low endotoxin levels, making it suitable for sensitive in vivo applications.
• No Cross-Reactivity: 55R-170 does not cross-react with TNFR2, ensuring specificity in experimental systems.

Summary Table

Clone 55R-170 remains a cornerstone tool in mouse immunology research, enabling precise dissection of TNFR1’s role in health and disease, and supporting the development of targeted therapeutic strategies.

Dosing regimens for clone 55R-170 (an anti-mouse TNFR1 monoclonal antibody) commonly fall within the range of 100–250 µg per mouse per dose in published in vivo protocols, typically delivered by intraperitoneal injection. However, there is no universally established standard regimen; dosing is often empirically adjusted based on the goals and disease context of each individual mouse model.

Key details and variations:

• The most frequently reported single dose is 100–250 µg per mouse for functional in vivo neutralization, similar to regimens used for other cytokine receptor-blocking antibodies.
• Dosing frequency and schedule (e.g., single vs. repeated doses, intervals) are highly variable and should be tailored to:
  • The mouse strain and disease model (e.g., acute vs. chronic inflammation, infection, cancer).
  • The expected pharmacokinetics and antibody half-life.
  • Endpoints of interest (e.g., acute cytokine blockade vs. sustained signaling inhibition).
• In vitro, 25 µg/mL of 55R-170 can inhibit about 50% of the biological effects of 1 ng/mL TNFα on L929 cells, but these concentrations do not directly translate to in vivo dosing.

Adaptation to mouse models:

• No compelling evidence suggests substantial differences in basic dosing regimen between genetic backgrounds (e.g., C57BL/6 vs. BALB/c) or major disease models, but specific experimental objectives (such as degree and duration of TNF signaling blockade) often prompt empirical adjustments.
• Some studies follow standard antibody dosing regimens used with other blocking antibodies in immunological mouse models—generally 100–250 µg/dose, administered as needed based on the disease course or experimental design.

Summary Table: Typical Clone 55R-170 Dosing in Mouse Models

Adjustments are often required depending on experimental design, and consulting primary literature for closely related applications is advised.

There is no evidence in the provided sources for radically different dosing based on mouse strain; rather, dosing is typically model- and hypothesis-driven.