The sequence of the first five N-terminal amino acids was determined and was found to be Met-Val-Arg-Ser-Ser.
The ED50 as determined by the cytolysis of murine L929 cells in the presence of Actinomycin D is < 0.05ng/ml, corresponding to a Specific Activity of 20MIU/mg.
1mg of TNF-alpha Human contain 20mM PBS, pH-7.2, and 10mM NaCl.
Sterile Filtered White lyophilized (freeze-dried) powder.
Protein quantitation was carried out by two independent methods
1. UV spectroscopy at 280 nm using the absorbency value of 1.234 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-a as a Reference Standard.
Greater than 98.0% as determined by
(a) Analysis by RP-HPLC.
(b) Analysis by SDS-PAGE.
It is recommended to reconstitute the lyophilized Tumor Necrosis Factor-alpha in sterile 18MΩ-cm H2O not less than 100ug/ml, which can then be further diluted to other aqueous solutions.
Lyophilized Tumor Necrosis Factor-a although stable at room temperature for 3 weeks, should be stored desiccated below -18oC. Upon reconstitution TNF-a should be stored at 4oC between 2-7 days and for future use below -18oC.
For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
Please prevent freeze-thaw cycles.
TNF-alpha, Tumor necrosis factor ligand superfamily member 2, TNF-a, Cachectin, DIF, TNFA, TNFSF2.
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