Thermo Fisher Scientific Rat TNF-alpha Recombinant Protein

Applications

• Western Blot (WB): Assay-dependent
• ELISA (ELISA): Assay-dependent
• Immunoprecipitation (IP): Assay-dependent
• Control (Ctrl): Assay-dependent

Product Specifications

Product Specific Information

The mature, soluble form of rat TNF-alpha was expressed in Pichia pastoris strain KM71H (GenBank Accession # P16599, amino acids 80–235).
Protein verification was performed by N-terminal sequencing, showing the first 10 amino acids as EAEALRSSSQ. The EAEA portion indicates incomplete cleavage of the alpha-Factor signal by Ste13 protease.
Purity is >95% by SDS-PAGE analyses.

Activity:
The biological activity of recombinant rat TNF-alpha was measured by its cytolytic effect on mouse WEHI-13VAR cells in the presence of Actinomycin D. The ED50 (or 1 unit of activity) for this effect is typically <5 pg/mL. For in vitro applications, a dose-response assay should be performed to determine the optimal concentration for specific experiments.

Reconstitution and Dilution:

• Store lyophilized material at 4°C.
• Reconstitute in 0.5 mL distilled or deionized water.
• Dilute in a buffer containing carrier protein, aliquot, and store at or below -70°C.
• Avoid multiple freeze-thaw cycles.
• Suitable buffers: PBS with 1% BSA or neutral media containing <10% FCS.

Target Information

TNF-alpha is a multifunctional proinflammatory cytokine belonging to the tumor necrosis factor (TNF) superfamily. It is primarily secreted by macrophages and binds to receptors TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR.
Functions include regulation of immune cells, cell proliferation, differentiation, apoptosis, lipid metabolism, and coagulation. TNF-alpha typically forms multimers and appears as a single 17 kDa band under non-reducing SDS-PAGE conditions.

Knockout studies in mice suggest a neuroprotective function, and TNF-alpha can cause tumor necrosis when injected into tumor-bearing mice. It also plays roles in immune responses to bacterial, viral, parasitic, and fungal infections, as well as in tumor cytotoxicity (synergistic with interferon-gamma).
This cytokine has been implicated in autoimmune diseases, insulin resistance, and cancer.

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