Anti-Mouse CD120b (TNFR2) [Clone TR75-54.7] — Purified in vivo GOLD™ Functional Grade

The clone TR75-54.7 is a monoclonal antibody that targets mouse Tumor Necrosis Factor Receptor Type II (TNFR2), also known as CD120b or p75. This antibody is used in in vivo mouse studies primarily for its ability to modulate TNFR2 signaling, which is involved in various biological processes including immunoregulation, cell proliferation, and apoptosis.

Applications of TR75-54.7 in In Vivo Studies:

1. Anti-Tumor Immunity: TR75-54.7 has been shown to enhance anti-tumor immunity when used in immuno-competent mice. It acts as a TNFR2 agonist when cross-linked in vitro, which can influence immune responses beneficially in the context of cancer.

2. Antitumor Effects: In mouse models of breast cancer, such as the 4T1 model, TR75-54.7 has demonstrated potent antitumor effects. It blocks TNFR2 signaling, which can promote tumor growth by supporting the suppressive function of T regulatory cells (Tregs).

3. Combination Therapy: The antibody is often used in combination with other treatments, such as CD25 antagonistic antibodies like PC61, to enhance its antitumor efficacy. This combination can lead to a more effective inhibition of tumor growth compared to monotherapies.

4. Immune Regulation: Given that TNFR2 is involved in immunoregulation, TR75-54.7 can be used to study the role of TNFR2 in immune responses and its potential as a therapeutic target in autoimmune diseases or cancer.

Overall, TR75-54.7 is a valuable tool in in vivo mouse studies for exploring the functions of TNFR2 and its potential applications in cancer immunotherapy.

The correct storage temperature for the sterile packaged clone TR75-54.7 (Anti-Mouse CD120b/TNFR2) is between 2°C and 8°C. It should be stored in this refrigerated condition without freezing and handled under aseptic conditions to maintain its integrity. For longer-term storage beyond one month, it is recommended to aseptically aliquot the solution into working volumes without diluting and store at -80°C, avoiding repeated freeze-thaw cycles.

Commonly Used Antibodies and Proteins with TR75-54.7

TR75-54.7 is a monoclonal antibody targeting mouse TNFR2 (CD120b), often used in research for flow cytometry, ELISA, immunoprecipitation, Western blotting, and receptor blocking. Its applications frequently involve pairing with other antibodies or proteins to maximize detection sensitivity, enable multiplex assays, or achieve functional outcomes.

Key Pairings in Experimental Workflows

• Biotinylated TR75-32.4: For ELISA setups, TR75-54.7 is commonly used as the capture antibody paired with biotinylated TR75-32.4 as the detection antibody. This combination allows for sensitive and specific detection of TNFR2 in sandwich ELISA formats.
• Biotinylated anti-Armenian Hamster IgG and Streptavidin-PE: In flow cytometry, to amplify signal detection when using TR75-54.7, a three-layer staining approach is recommended. This involves using TR75-54.7 as the primary antibody, followed by a biotinylated anti-Armenian hamster IgG secondary antibody, and finally streptavidin-phycoerythrin (SAv-PE) for fluorescence detection.
• PC61 (anti-CD25): In functional in vivo studies, especially in cancer immunotherapy research, TR75-54.7 has been co-administered with PC61, a monoclonal antibody targeting CD25 (IL-2 receptor ? chain). The combination of TR75-54.7 and PC61 showed superior antitumor effects compared to either antibody alone in a mouse breast cancer model, highlighting their utility in combinatorial blockade strategies targeting regulatory T cell subsets.

Additional Notes on Companion Proteins

• Recombinant Mouse TNF R2: TR75-54.7 itself is raised against purified recombinant mouse TNF R2, which is also used for immunization and as a positive control in various assays.
• Carrier Proteins: The antibody is typically formulated in phosphate-buffered saline (PBS) without carrier proteins (e.g., BSA) or preservatives for in vivo applications, but BSA or other blockers may be added in assay buffers for specific applications.

Summary Table: Common Antibody/Protein Pairings with TR75-54.7

Conclusion

TR75-54.7 is frequently used alongside biotinylated TR75-32.4 for ELISA, with secondary detection systems in flow cytometry, and in combination with PC61 for in vivo functional studies. These pairings are standard in the literature for TNFR2 research, enabling sensitive detection, multiplex analysis, and combinatorial therapeutic strategies.

Clone TR75-54.7 is a widely used hamster monoclonal antibody targeting mouse TNF receptor 2 (TNFR2, also known as CD120b or p75), and it has been extensively cited in scientific literature for its applications in immunology and cancer research.

Key findings from citations using TR75-54.7:

• Agonist and Competitive Activity: TR75-54.7 competes with TNF-? for binding to TNFR2 and can function as a TNFR2 agonist in vitro when cross-linked, leading to proliferation of specific T cell lines and activation of the NF-?B signaling pathway. Bio-layer interferometry and cell-based reporter assays confirm its role as an agonist and its ability to mimic the effects of TNF-? on TNFR2.

• Antitumor Immune Modulation: In immunocompetent mouse models, TR75-54.7 and related anti-TNFR2 antibodies have been shown to enhance anti-tumor immunity by altering the tumor microenvironment. Specifically, TR75-54.7 treatment leads to significant increases in CD8+ T cell numbers and a reduction of immunosuppressive regulatory T cell (Treg) activity in tumors.

• Targeting Treg Cells: TR75-54.7-mediated TNFR2 blockade downregulates markers of activation (e.g., CTLA4, Tnfrsf1b) in Tregs, especially the activated subset (aTregs). This not only decreases their suppressive phenotype but also reduces their clonal expansion, fostering a less suppressive (more immune-permissive) tumor environment.

• Mechanism of Action and Biological Effects:

  • Blocks ligand-induced receptor signaling: TR75-54.7 effectively blocks TNF-? or LT-? induced signaling through TNFR2, affecting downstream processes such as apoptosis, NF-?B activation, inflammation, and immune cell regulation.
  • Fc?R Interaction: The antibody is cross-linked in vivo mainly via mouse Fc?RII and Fc?RIII, but does not mediate antibody-dependent cellular phagocytosis (ADCP).

• Experimental Utility and Applications:

  • Used for flow cytometry, blocking experiments, immunoprecipitation, ELISA, and functional studies of TNFR2 biology.
  • Frequently cited in studies interrogating the role of TNFR2 in immune regulation, cancer immunotherapy, and inflammation.

Summary Table: Functional Findings for TR75-54.7

Overall, clone TR75-54.7 is considered a reliable tool for dissecting the roles of TNFR2 in the mouse immune system, and its use is associated with discoveries regarding immune activation, antitumor responses, and the biology of Tregs in cancer and inflammation.