| TNF is secreted by macrophages, monocytes, neutrophils, T-cells, NK-cells following their stimulation by bacterial LPS. Cells expressing CD4 secrete TNF-alpha while CD8 cells secrete little or no TNF-alpha. The synthesis of TNF-alpha is induced by many different stimuli including interferons, IL2, GM-CSF. |
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| Catalog # | T9160-10A |
| Recombinant Human Tumor Necrosis Factor-alpha is a single, non-glycosylated, polypeptide chain containing 158 amino acids and having a molecular mass of 17483.77 Dalton. |
| Sequence | The sequence of the first five N-terminal amino acids was determined and was found to be Met-Val-Arg-Ser-Ser. |
| Dimers and Aggregates | 1% as determined by silver-stained SDS-PAGE gel analysis. |
| Biological Activity | Recombinant Tumor Necrosis Factor-alpha is fully biologically active when compared to standard. The ED50 as determined by the cytolysis of murine L929 cells in the presence of Actinomycin D is less then 0.05ng/ml, corresponding to a Specific Activity of 2 x 10e7 IU/mg. |
| Protein Content | Protein quantitation was carried out by two independent methods: |
| 1. UV spectroscopy at 280 nm using the absorbency value of 1.24 as the extinction coefficient for a 0.1% (1mg/ml) solution. This value is calculated by the PC GENE computer analysis program of protein sequences (IntelliGenetics). |
| 2. Analysis by RP-HPLC, using a calibrated solution of TNF-alpha as a Reference Standard. |
| Storage and Stability | Lyophilized powder may be stored at 4°C for short-term only. Reconstitute to nominal volume by adding sterile ddH2O, 0.1% HSA or BSA and store at -20°C. Reconstituted product is stable for 12 months at -20°C. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made in assay buffer. |
| Source | Recombinant, Human (E. coli) |
| Purity | 95% by RP-HPLC, FPLC, or reducing/non-reducing SDS-PAGE Silver Stain. Chromatographically purified. Endotoxin: <0.1ng/ug (IEU/ug). |
| Concentration | ~0.1mg/ml |
| Form | Supplied as a liquid in 20mM PBS, pH 7.2, 0.1% BSA. |
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| Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
