Technical Data
Tumor Necrosis Factor alpha, Recombinant, Rat (TNFa)
Molecular Biology Storage: -20CShipping: Blue Ice
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.

Recombinant 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 Sequence:
The sequence of the first five N-terminal amino acids was determined and was found to be Met-Leu-Arg-Ser-Ser.

Dimers and Aggregates:
Less than 1% as determined by silver-stained SDS-PAGE gel analysis.

Biological Activity:
Rat 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.

0.1ng/ug (IEU/ug) of Rat TNF-alpha.

Protein Content:
Protein 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.

It is recommended to reconstitute the lyophilized Rat TNF-alpha in sterile 18M -cm H2O not less than 100g/ml, which can then be further diluted to other aqueous solutions.

Lyophilized 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 Weight:
Source: E. coli
Purity: 97% as determined by:
(a) Analysis by RP-HPLC.
(b) Anion-exchange FPLC.
(c) Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel.
Concentration: As reported
Form: Supplied as a lyophilized powder without additives.

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.
1. Who might be predisposed to the development of serious side effects when treated with TNF-alpha antagonist?
Clin Exp Rheumatol 2006 Mar-Apr;24(2):211; author reply 211
2. The effect of periodontal therapy on TNF-alpha, IL-6 and metabolic control in type 2 diabetics.
J Dent Hyg 2006 Spring;80(2):7
3. Cell to cell contact through ICAM-1-LFA-1 and TNF-alpha synergistically contributes to GM-CSF and subsequent cytokine synthesis in DBA/2 mice induced by 1,3-beta-D-Glucan SCG.
J Interferon Cytokine Res 2006 Apr;26(4):235-47
4. TNF-alpha is necessary for induction of coronary artery inflammation and aneurysm formation in an animal model of Kawasaki disease.
J Immunol 2006 May 15;176(10):6294-301
5. Interleukin-17 acts i ndependently of TNF-alpha under arthritic conditions.
J Immunol 2006 May 15;176(10):6262-9
6. Peptide YY attenuates STAT1 and STAT3 activation induced by TNF-alpha in acinar cell line AR42J.
J Am Coll Surg 2006 May;202(5):788-96

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.