- TNF is secreted by macrophages, monocytes, neutrophils, T-cells, NK-cells following their stimulation by bacterial LPS. The synthesis of TNF-α is induced by many different stimuli including interferons, IL2 and GM-CSF. It is a potent lymphoid factor which exerts cytotoxic effects on a wide range of tumor cells and certain other target cells. The clinical use of the potent anti-tumor activity of TNF-α has been limited by the proinflammatory side effects including fever, dose-limiting hypotension, hepatotoxicity, intravascular thrombosis and hemorrhage. Thus, designing clinically applicable TNF-α mutants with low systemic toxicity has been of great pharmacological interest. Human TNF-α, which only binds to the murine TNF-R55, exhibits retained anti-tumor activity and reduced systemic toxicity in mice compared with the murine TNF-α. Based on these results, many TNF-α 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. Compared with the wild-type, recombinant human TNF-alpha Variant/Mutant has an amino acid sequence deletion from a.a. 1-7, and the following a.a. substitutes Arg8, Lys9, Arg10 and Phe157. It is proven to have more activity and less inflammatory side effects in vivo. Recombinant Human TNF-α Variant produced in E. coli is a single, non-glycosylated polypeptide chain of 151 amino acids and a molecular mass of 16598 Dalton.
- Product Overview
- Human TNF expressed in E.coli
- CAT# CSC-CTK0346-50 (50 μg); CAT# CSC-CTK0346-250 (250 μg)
- Greater than 95% as determined by SDS-PAGE, FPLC and RP-HPLC analysis.
- Human TNF-alpha M Variant is fully biologically active when compared to the wild type. 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 1 x 10^8 IU/mg.
- Endotoxin Level
- Less than 1 EU/μg.
- Sterile-filtered, white, lyophilized (freeze-dried) powder. The protein was lyophilized after extensive dialysis against 0.5x PBS (pH 7).
- Please centrifuge product briefly before opening the vial. The lyophilized rHu TNF- alpha M should be reconstituted in sterile, ultra-pure water to a concentration of 0.1-1.0 mg/ml. This solution can then be diluted into other aqueous buffers and stored at -20°C for future use.
- Storage & Stability
- The lyophilized protein, though stable at room temperature for up to 3 weeks, is best stored desiccated at -20°C. Reconstituted rHu TNF-alpha M should be used immediately or stored long- term in undiluted working aliquots at -20°C. For long-term storage it is recommended to add a carrier protein (0.1% HSA or BSA). Avoid repeated freeze-thaw cycles.
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