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You are here:Home » Growth Factors, Cytokines » Growth Factors-TNF » Tumor Necrosis Factor alpha, Recombinant, Rat (TNFa)

Tumor Necrosis Factor alpha, Recombinant, Rat (TNFa)

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Specifications

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.
Catalog #T9160-20B
DescriptionRecombinant rat TNF-alpha produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 157 amino acids and having a molecular mass of 17339.44 Dalton. Recombinant Rat TNF-alpha is purified by standard chromatographic techniques.
Amino Acid SequenceThe sequence of the first five N-terminal amino acids was determined and was found to be Met-Leu-Arg-Ser-Ser.
Dimers and AggregatesLess than 1% as determined by silver-stained SDS-PAGE gel analysis.
Biological ActivityRat TNF-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 < 0.05ng/ml, corresponding to a Specific Activity of 5x107 IU/mg.
Endotoxin 0.1ng/ug (IEU/ug) of Rat TNF-alpha.
Protein ContentProtein quantitation was carried out by two independent methods:
1. UV spectroscopy at 280 nm using the absorbency value of 1.25 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.
SolubilityIt is recommended to reconstitute the lyophilized Rat TNF-alpha in sterile 18M -cm H2O not less than 100µg/ml, which can then be further diluted to other aqueous solutions.
StabilityLyophilized Rat TNF-alpha although stable at room temperature for 3 weeks, should be stored desiccated below -180C. Upon reconstitution Rat TNF-alpha should be stored at 40C between 2-7 days and for future use below -180C. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA). Please prevent freeze-thaw cycles.
Molecular Weight17.34kD
SourceE. coli
Purity97% as determined by:
(a) Analysis by RP-HPLC.
(b) Anion-exchange FPLC.
(c) Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel.
ConcentrationAs reported
FormSupplied as a lyophilized powder without additives.
Important NoteThis 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.


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