Technical Data
T9160-10M
Tumor Necrosis Factor alpha, Mutant, Recombinant, Human (TNFa)
10ug
50ug
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.

Description:
The clinical use of the potent anti-tumor activity of TNF-alfa has been limited by the proinflammatory side effects including fever, dose-limiting hypotension, hepatotoxicity, intravascular thrombosis, and hemorrhage. Designing clinically applicable TNF-a mutants with low systemic toxicity has been an intense pharmacological interest. Human TNF- , which binds to the murine TNF-R55 but not to the mouse TNF-R75, exhibits retained anti-tumor activity and reduced systemic toxicity in mice compared with murine TNF-a, which binds to both murine TNF receptors. Based on these results, many TNF- mutants that selectively bind to TNF-R55 have been designed. These mutants displayed cytotoxic activities on tumor cell lines in vitro, and exhibited lower systemic toxicity in vivo. Recombinant Human TNF-alfa Variant/Mutant compared with the wild-type, has an amino acid sequence deletion from a.a. 1-7, and the following a.a. substitutes Arg8, Lys9, Arg10 and Phe157 which is proven to have more activity and with less inflammatory side effect in vivo. Recombinant Human TNF-a Variant produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 151 amino acids and having a molecular mass of 16598 Dalton. The Human TNF-alpha Variant is purified by standard chromatographic techniques.

AA Sequence:
MRKRKPVAHV VANPQAEGQL QWLNRRANAL LANGVELRDN QLVVPSEGLY LIYSQVLFKG QGCPSTHVLL THTISRIAVS YQTKVNLLSA IKSPCQRETP EGAEAKPWYE PIYLGGVFQL EKGDRLSAEI NRPDYLDFAE SGQVYFGIIAF.

Biological Activity:
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 2.0x10e7 U/mg.

Storage and Stability:
Lyophilized powder may be stored at -20C. Stable for 12 months at -20C. Reconstitute with sterile ddH2O, 0.1% HSA or BSA. Aliquot to avoid repeated freezing and thawing. Store at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

Molecular Weight:
16.6kD
Source: E. coli
Purity: 95% as determined by RP-HPLC, anion-exchange FPLC and/or reducing and non-reducing SDS-PAGE Silver Stained gel. Endotoxin: 0.1ng/ug (IEU/ug))
Concentration: As reported
Form: Supplied as a lyophilized powder in 0.5X PBS pH 7.0. Reconstitute with sterile dH2O to 0.1mg/ml.

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.