Mouse TNFα Antibody – Purified in vivo GOLD™ Functional Grade

Clone TN3-19.12 is used in in vivo mouse studies primarily for neutralizing murine TNF-? (tumor necrosis factor-alpha) and cross-reacts with mouse TNF-?. This antibody is crucial for understanding the role of TNF in various biological processes, including inflammation and immune responses.

Applications of TN3-19.12 in Mouse Studies:

1. Neutralization of TNF-? in Endotoxin Shock:

   • TN3-19.12 has been shown to effectively neutralize endogenously produced TNF in mice, preventing its detection in circulation and protecting mice from the lethal effects of endotoxin shock when injected before LPS (lipopolysaccharide) treatment.

2. Investigation of TNF-? and TNF-? Roles:

   • By neutralizing TNF-?, researchers can study its impact on disease models, such as autoimmunity and inflammation, and discern the roles of TNF-? versus TNF-? in these processes.

3. In Vivo Models of Disease:

   • This antibody is used in in vivo models to study the effects of TNF-? blockade on disease progression, such as in models of arthritis or other inflammatory conditions.

4. Dosage and Administration:

   • While specific dosages may vary based on the study design, the antibody is typically administered via intraperitoneal injection to ensure systemic exposure.

In summary, TN3-19.12 is a valuable tool for in vivo studies, enabling researchers to explore the biological functions of TNF-? and TNF-? and their implications in various disease contexts. Its use helps to elucidate the mechanisms underlying inflammatory responses and immune regulation in mice.

The correct storage temperature for the sterile packaged clone TN3-19.12, which is a mouse TNF-alpha antibody, depends on the duration of storage:

• Short-term storage (up to one month): Store at 2 to 8°C under sterile conditions.
• Long-term storage: Store at ? -70°C after aseptically aliquoting into working volumes. Alternatively, storage at -20°C to -80°C is recommended for maintaining stability and reducing microbial growth.

The TN3-19.12 antibody is commonly paired with several other antibodies and proteins in various experimental applications, particularly for detecting and studying TNF-? in research settings.

Detection Antibodies for ELISA and ELISPOT Assays

The most frequently paired detection antibody with TN3-19.12 is the biotinylated Poly5160 antibody. This combination forms the foundation of sandwich ELISA and ELISPOT assays for mouse/rat TNF-? detection. In these assays, TN3-19.12 serves as the capture antibody at concentrations of 2-6 µg/ml, while the biotinylated Poly5160 functions as the detecting antibody.

Comparative Antibodies in Research

In validation studies and comparative analyses, researchers have used polyvalent rabbit anti-rTNF antibodies alongside TN3-19.12. These rabbit antibodies have been employed to establish TNF-specific ELISA systems and to confirm the specificity and neutralizing capacity of TN3-19.12. The combination of monoclonal TN3-19.12 and polyclonal rabbit anti-TNF antibodies has proven effective in creating highly sensitive detection systems capable of measuring picogram quantities of TNF.

Cross-Reactive Recognition Targets

TN3-19.12 demonstrates cross-reactivity with multiple TNF family members, which makes it useful in studies examining both TNF-? and TNF-? (lymphotoxin). Research has shown that this antibody recognizes both forms, with TNF-? appearing as a 17 kDa band and TNF-? as a glycosylated 24.7 kDa form in Western blot analyses. This dual recognition capability makes TN3-19.12 particularly valuable in studies examining T cell-derived cytokines, where both TNF-? and TNF-? are commonly expressed.

Endotoxin Studies and In Vivo Applications

In endotoxin shock studies, TN3-19.12 has been used in conjunction with lipopolysaccharide (LPS) treatments to demonstrate its neutralizing capabilities. These studies have shown that TN3-19.12 can effectively prevent TNF detection in circulation and protect mice from lethal endotoxin effects when administered before LPS challenge.

The antibody's versatility in multiple experimental contexts, from basic sandwich immunoassays to complex in vivo neutralization studies, has made it a cornerstone reagent in TNF research, particularly when combined with these complementary detection and validation tools.

Key Findings on Clone TN3-19.12 in Scientific Literature

Neutralization of Murine TNF Activity

• The monoclonal antibody from clone TN3-19.12, purified from Armenian hamster immunization, neutralizes 100% of the lytic activity of both recombinant and natural murine tumor necrosis factor-alpha (MuTNF-?) at a specific antibody input (25 ng per unit).
• The antibody also inhibits all lytic activity in culture supernatants from various T cell sources, including activated T cell clones and hybridomas that express high levels of TNF-? and TNF-? (lymphotoxin, LT).
• In vivo, injection of TN3-19.12 into mice prevented detection of TNF in circulation and protected mice from the lethal effects of endotoxin shock, demonstrating effective neutralization of endogenously produced TNF.
• The antibody is also used in commercial contexts for neutralization studies, particularly in mouse and rat models.

Cross-reactivity and Molecular Recognition

• Western blot analysis demonstrated that TN3-19.12 recognizes both the 17 kDa MuTNF-? and the 24.7 kDa, glycosylated TNF-? (LT), showing cross-reactivity between these two cytokines.
• Commercial descriptions confirm that TN3-19.12 neutralizes murine TNF-? and cross-reacts with mouse TNF-?.

ELISA Development and Bioanalytical Applications

• TN3-19.12 and polyvalent rabbit anti-rTNF were used to develop a MuTNF-specific ELISA capable of detecting picogram quantities of TNF, both recombinant and natural.
• Peak TNF serum levels (11 ng/ml) were detected in mice challenged with a lethal dose of LPS, declining rapidly thereafter.

Paradoxical Effects in Infectious Disease Models

• In Trypanosoma cruzi-infected mice, administration of anti-TNF monoclonal antibody TN3 (presumably TN3-19.12) led to increased rather than decreased plasma TNF levels, worse cachexia, and higher mortality.
• This effect is thought to result from prolonged TNF half-life due to complexation with the antibody, leading to higher amounts of free TNF late in infection.
• The paradox was not due to endotoxin contamination and did not occur in noninfected animals, suggesting a disease-specific phenomenon.

Summary Table

Expert Interpretation

TN3-19.12 is a highly effective neutralizing antibody for murine TNF-?, with cross-reactivity to TNF-?, and has been instrumental in both basic research and assay development. Its in vivo use in healthy models demonstrates potent TNF inhibition and protective effects, but in certain disease contexts (e.g., T. cruzi infection), it may paradoxically exacerbate pathology by altering TNF kinetics and availability. These findings underscore the importance of context when interpreting the effects of anti-TNF therapies in experimental models.