Contents: Recombinant Human Tumor Necrosis Factor alpha (TNF-α)
Catalog Number: 14-8329
Formulation: Sterile liquid; 10 mM sodium phosphate, pH 7.2, 150 mM NaCl, with 0.5% BSA. 0.22 µm filtered.
Storage Conditions: For greatest stability, keep concentration of primary stock at or above 10 µg/ml. For long term storage, aliquot into polypropylene vials (volumes of 20 µl or greater) and store at or below -80°C. Avoid repeated freeze/thaw cycles.
Handling Conditions: For best recovery, always quick-spin vial prior to opening. For dilution of current stock, always include carrier protein (1% BSA or 10% FBS) in the buffered saline diluent.
Source: E. coli expressed amino acids Val 77-Leu 233 of human TNF-α (accession # HUMTNFAB).
Molecular Mass: The protein is not methionylated at the N-terminal and has a predicted molecular mass of 17,352. The DTT-reduced and non-reduced protein migrate as a 16 kDa polypeptides on SDS-PAGE. The native protein is greater than 95% homotrimer as determined by crosslinking with disuccinimidyl suberate and by size exclusion chromatography.
Purity: Greater than 98%, as determined by SDS-PAGE
Endotoxin Level: Less than 0.01 ng/ug cytokine as determined by the LAL assay.
Bioactivity: Measured by mouse L929 cytotoxicity assay. The ED50 is 0.01 ng/ml, corresponding to a specific activity of 1.0 x 108 Units/mg.
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Description
Human TNF-α is a 17 kDa factor produced by macrophages, monocytes, neutrophils, CD4
+ T cells and NK cells. A 26 kDa form of TNF-α is expressed as a membrane bound molecule. TNF-α is cytolytic and plays an important role in immune regulation. Dimers and trimers of TNF-α have been observed.
Applications Reported
For research use only, not for diagnostic or therapeutic use. Recombinant human TNF-α is biologically active and can induce cytolysis of L929 mouse cell line in culture. It is also used as cytokine in human TNF-α ELISA.
Applications Tested
This recombinant human TNF-α has been tested by sandwich ELISA using anti-human TNF-α antibody pair (MAb1/MAb11) and in bioassay using mouse cell line L929. The ED
50 measured in a L929 killing assay is typically 0.01 ng/ml, corresponding to a specific activity of approximately 1.0 x10
8 Units/mg.
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