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Synonyms
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Tumor Necrosis Factor, TNFSF2, Cachectin, Differentiation-inducing factor (DIF), Necrosin, Cytotoxin
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Species
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Human
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Accession
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P01375
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GeneID
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7124
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Source
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Escherichia coli.
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Molecular Weight
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重组人类肿瘤坏死因子-α/ TNFSF2，变体蛋白Approximately 16.9 kDa, a single non-glycosylated polypeptide chain containing 151 amino acids. Compared with the wild-type, rHuTNF-α Variant has an amino acid sequence (a.a.) 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.
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Quantity
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10µg/50µg/1000µg
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AA Sequence
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MRKRKPVAHV VANPQAEGQL QWLNRRANAL LANGVELRDN QLVVPSEGLY LIYSQVLFKG QGCPSTHVLL THTISRIAVS YQTKVNLLSA IKSPCQRETP EGAEAKPWYE PIYLGGVFQL EKGDRLSAEI NRPDYLDFAE SGQVYFGIIA F
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Purity
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> 98 % by SDS-PAGE and HPLC analyses.
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Biological Activity
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重组人类肿瘤坏死因子-α/ TNFSF2，变体蛋白Fully biologically active when compared to standard. The ED50 as determined by a cytotoxicity assay using murine L929 cells is less than 0.01 ng/ml, corresponding to a specific activity of > 1.0 × 107 IU/mg in the presence of actinomycin D.
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Physical Appearance
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Sterile Filtered White lyophilized (freeze-dried) powder.
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Formulation
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Lyophilized from a 0.2 μm filtered concentrated solution in PBS, pH 7.0.
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Endotoxin
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Less than 1 EU/μg of rHu TNF-α/TNFSF2, Variant as determined by LAL method.
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Reconstitution
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重组人类肿瘤坏死因子-α/ TNFSF2，变体蛋白We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Reconstitute in sterile distilled water or aqueous buffer containing 0.1 % BSA to a concentration of 0.1-1.0 mg/mL. Stock solutions should be apportioned into working aliquots and stored at ≤ -20 °C. Further dilutions should be made in appropriate buffered solutions.
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Storage
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This lyophilized preparation is stable at 2-8 °C, but should be kept at -20 °C for long term storage, preferably desiccated. Upon reconstitution, the preparation is stable for up to one week at 2-8 °C. For maximal stability, apportion the reconstituted preparation into working aliquots and store at -20 °C to -70 °C. Avoid repeated freeze/thaw cycles.
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重组人类肿瘤坏死因子-α/ TNFSF2，变体蛋白
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SDS-PAGE
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Reference
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1. Davenport C, Kenny H, Ashley DT, et al. 2012. Eur J Clin Invest, 42: 1173-9.
2. Cavalcanti YV, Brelaz MC, Neves JK, et al. 2012. Pulm Med, 2012: 745483.
3. Sheng WS, Hu S, Ni HT, et al. 2005. J Leukoc Biol, 78: 1233-41.
4. Berthold-Losleben MandHimmerich H. 2008. Curr Neuropharmacol, 6: 193-202.
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Background
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The clinical use of the potent antitumor activity of TNF-αhas been limited by the proinflammatory side effects including fever, dose-limiting hypotension, hepatotoxicity, intravascular thrombosis, and hemorrhage. Designing clinically applicable TNF-αmutants with low systemic toxicity has been an intense pharmacological interest. Human TNF-α, which binds to the murine TNF-R55 but not to the murine TNF-R75, exhibits retained antitumor activity and reduced systemic toxicity in mice compared with murine TNF-α, which binds to both murine TNF receptors. Based on these results, many TNF-αmutants 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.
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