Anti-Mouse TNFα [Clone TN3-19.12] — Purified in vivo PLATINUM™ Functional Grade

In Vivo Applications of TN3-19.12 in Mice

Clone TN3-19.12 is a hamster monoclonal antibody developed against murine TNF-α, widely used in mouse research for its potent neutralizing activity and cross-reactivity with mouse TNF-β (lymphotoxin, LT). Its common in vivo applications include:

Key Applications

• TNF-α Neutralization: The primary use of TN3-19.12 is to neutralize endogenous TNF-α (and to some extent TNF-β) in mouse models, allowing researchers to dissect the physiological and pathological roles of TNF in immune responses, inflammation, and disease pathogenesis. For example, administering TN3-19.12 prior to LPS challenge prevents detection of TNF in circulation and protects mice from lethal endotoxin shock, demonstrating its efficacy in blocking endogenous TNF activity.
• Immune and Inflammatory Disease Models: TN3-19.12 is used in various disease models (e.g., autoimmune diseases, inflammatory conditions, and septic shock) to determine the contribution of TNF to disease progression and to test therapeutic interventions.
• In Vivo Cytokine Capture Assay (IVCCA): TN3-19.12 serves as the capture antibody in the IVCCA, a method that dramatically increases the in vivo half-life and detection sensitivity of cytokines in serum, facilitating measurement of cytokine production during immune responses. This assay is especially useful for tracking cytokine dynamics in real time in living animals.
• Functional Studies of Cytokine Networks: By neutralizing TNF-α, researchers can investigate its role in cytokine networks, cell signaling, and cross-regulation with other cytokines in vivo.

Technical Notes

• Dosing: A typical in vivo dose is 250 µg per mouse, though optimal dosing may vary depending on the experimental model.
• Specificity: TN3-19.12 neutralizes both TNF-α and TNF-β, making it a broad tool for blocking TNF family cytokines in mice.
• Formulation: The antibody is available in low-endotoxin, functional-grade preparations suitable for in vivo use, minimizing the risk of confounding immune responses.
• Cross-Reactivity: While primarily used in mice, TN3-19.12 also cross-reacts with rat and rabbit TNF-α, expanding its utility to other species.

Conclusion

TN3-19.12 is a versatile tool for in vivo neutralization of TNF in mice, with applications spanning basic immunology, disease modeling, and cytokine biology. Its ability to block both TNF-α and TNF-β, coupled with its use in advanced assays like the IVCCA, makes it a mainstay in inflammation and immune research.

The TN3-19.12 monoclonal antibody, which targets mouse, rat, and rabbit TNFα, is often used in combination with several other antibodies and proteins, particularly for immunoassays like ELISA, ELISPOT, and flow cytometry.

Commonly co-used antibodies and proteins with TN3-19.12 in the literature include:

• Biotinylated Poly5160 (anti-mouse/rat TNFα polyclonal antibody): Frequently used as the detecting antibody in sandwich ELISA and ELISPOT when TN3-19.12 serves as the capture antibody.
• Isotype controls: Utilized as negative controls to confirm the specificity of TN3-19.12's binding in flow cytometry and other assays.
• Fluorochrome-conjugated secondary antibodies: Examples include antibodies conjugated to APC (Allophycocyanin) or other fluorophores for flow cytometric analysis and intracellular staining.
• Alternative capture/detection clones: Clone 1F3F3D4 is recommended as a capture antibody for ELISPOT applications, indicating that TN3-19.12 may be paired with this clone depending on assay requirements.
• Recombinant TNFα (mouse or rat): Used as a standard protein for quantitation in ELISA assays in combination with TN3-19.12.
• Irrelevant antibody controls (e.g., L2-3D9): Employed in animal model studies to validate TN3-19.12 specificity by comparison.

Other relevant notes:

• TN3-19.12 has been paired with polyvalent rabbit anti-rTNF and other monoclonal antibodies in characterization and specificity studies.
• It is occasionally used with anti-TNFβ (LT) reagents since TN3-19.12 may cross-react with mouse TNFβ (lymphotoxin).
• In general, selection of the co-antibody depends on the experimental format (e.g., flow cytometry, ELISA, in vivo neutralization), detection system (biotin, fluorophore), and the species involved.

These combinations allow researchers to achieve sensitive and specific detection, quantitation, or neutralization of TNFα in experimental models.

Clone TN3-19.12 is a widely cited monoclonal antibody notable for its ability to neutralize murine (mouse and rat) tumor necrosis factor-alpha (TNF-α), a key cytokine involved in inflammation and immune regulation.

Key findings from scientific literature using this clone include:

• Neutralization of TNF-α Bioactivity: TN3-19.12 effectively neutralizes both natural and recombinant TNF-α, making it a central tool for investigating the biological functions of TNF-α in inflammation, autoimmunity, and infection models.
• In Vivo and In Vitro Functional Use: The antibody is frequently used in mouse models to block TNF-α activity in vivo, as well as in vitro assays such as intracellular staining for flow cytometry, ELISA, and the In Vivo Cytokine Capture Assay (IVCCA), which greatly improves sensitivity for cytokine detection.
• Impact on Disease Models: In infectious disease models, administration of TN3-19.12 can paradoxically increase circulating TNF-α (likely by stabilizing the cytokine in immune complexes), alter sTNFR/TNF ratios, and exacerbate pathology, as observed in Trypanosoma cruzi infection—leading to accelerated cachexia and mortality. This underscores the complex dynamics of TNF-α neutralization in vivo.
• Mechanistic Studies: The antibody has been used to demonstrate the role of TNF-α in mobilizing immune cells (monocytes, neutrophils, macrophages, lymphocytes) and modulating cytokine cascades during acute and chronic inflammatory responses.
• Safety and Side Effects: Long-term anti-TNF-α treatment (TN3 and others) can be associated with increased risk of lymphomas in mouse models of autoimmunity, highlighting implications for chronic blockade of this pathway.
• Technical Features: TN3-19.12 is an Armenian hamster IgG, suitable for both rat and mouse TNF-α, and is recommended for use in functional blocking/neutralization assays and immune phenotyping via flow cytometry.
• Other Cross-Reactivity: The antibody may cross-react with murine TNF-β (also known as lymphotoxin-α), though it is primarily characterized for TNF-α targeting.

These findings position TN3-19.12 as a “gold standard” tool for dissecting TNF-α-driven mechanisms in immunology and disease research, with both methodological and experimental insights directly supported by its extensive use across the literature.

Dosing regimens of clone TN3-19.12—an anti-mouse TNF-α monoclonal antibody—vary significantly across mouse models, primarily depending on the disease context, route of administration, and study objectives.

Key dosing regimens:

• Acute neutralization in inflammatory models: Typical doses are 25–30 mg/kg intravenously (i.v.), administered around the time of an inflammatory or infectious insult.
• Chronic inflammation models (colitis, TRUC mice): A common regimen is 15 mg/kg intraperitoneally (i.p.) once weekly for 4 weeks.
• Fixed per-mouse doses (general in vivo use): Some studies use 250 µg per mouse i.p., particularly for acute neutralization studies or when a per-mouse basis is more appropriate.
• Autoimmunity/diabetes prevention (NOD mice): Dosing examples include 20 µg/g (i.e., 20 mg/kg) or 100 µg/injection i.p. every other day for 3–4 weeks.
• Endotoxin (LPS shock) models: Protective effects were demonstrated, but the explicit dosing was not detailed in every publication; often, doses in the 10–30 mg/kg range are reported or inferred.

Route of administration also varies: most studies utilize intraperitoneal (i.p.) or intravenous (i.v.) routes, with i.p. being the most common for routine in vivo neutralization.

Additional notes:

• The selection of dose and frequency is influenced by the disease model, desired duration of neutralization, and mouse size.
• Some sources recommend that investigators empirically titrate doses for their particular application, as performance and requirements can vary between models and endpoints.

Citations reflect best practices and reported regimens across common disease models; individual protocols sometimes require adjustment.