Thermo Fisher Scientific TNF alpha Monoclonal Antibody (TN3-19.12), Functional Grade, eBioscience

Thermo Fisher Scientific TNF alpha Monoclonal Antibody (TN3-19.12), Functional Grade, eBioscience

Applications and Tested Dilution

Product Specifications

Product Specific Information

The TN3-19.12 monoclonal antibody reacts with mouse, rat, and rabbit TNF alpha, but not with human TNF alpha. It is a neutralizing antibody. Mouse TNF alpha (17 kDa) is produced by macrophages, monocytes, neutrophils, CD4+ T cells, and NK cells; a 26 kDa membrane-bound form also exists. TNF alpha plays an important role in immune regulation and cytolytic activity.

Applications Reported:

• Intracellular staining (flow cytometry)
• Cytokine neutralization
• ELISA

Fluorochrome-conjugated TN3-19.12 is recommended for intracellular staining and flow cytometry. Functional Grade purified TN3-19.12 (Product # 16-7423) is recommended for functional assays. For ELISPOT capture, use clone 1F3F3D4 (Product # 16-7325).

Applications Tested:

• Verified low endotoxin level by LAL assay
• Tested for neutralization of TNF alpha bioactivity and ELISA capture

At 0.150 µg/mL, TN3-19.12 inhibits 50% of the biological effects of 1.0 ng/mL mouse TNF alpha in an L929 cytotoxicity assay. Protocols for cytokine bioassays and neutralization can be found in BestProtocols®.

ELISA Usage:

• Capture antibody for mouse/rat TNF alpha
• Combine with biotinylated polyclonal antibody (Product # 13-7341-81)
• Standard: recombinant mouse TNF alpha ([Product # 14-8321-62]) or rat TNF alpha
• Recommended concentration: 1–4 µg/mL
• Typical standard curve: 2000 pg/mL–15 pg/mL (mouse) or 5000–30 pg/mL (rat)

Recognition of mouse TNF alpha is not interfered by 1000× excess soluble TNF alpha receptor type I or II.

Additional Data:

• Filtration: 0.2 µm post-manufacturing filtered
• Purity: >90% (SDS-PAGE)
• Endotoxin Level: <0.001 ng/µg antibody (LAL assay)
• Aggregation: <10% (HPLC)
• Storage and Handling: Use in sterile environment

Target Information

TNF alpha is a multifunctional proinflammatory cytokine of the TNF superfamily. Secreted mainly by macrophages, it binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. TNF alpha regulates immune cells, proliferation, differentiation, apoptosis, lipid metabolism, and coagulation. It forms multimers (dimers, trimers, pentamers) and shows a 17 kDa band under non-reducing SDS-PAGE.

Knockout studies indicate neuroprotective functions and tumor necrosis activity. TNF alpha is involved in immune responses to infections and tumor cytolysis, synergistic with interferon-gamma. It is implicated in autoimmune diseases, insulin resistance, and cancer.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.