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Mouse TNF-α, amino acids Leu80-Leu235 (Accession # NM_013693), was expressed in E. coli.
The 156 amino acid recombinant protein has a predicted molecular mass of 17,257 Da. The DTT-reduced and non-reduced protein migrate at approximately 16 kD by SDS-PAGE. The N-terminal amino acid is Leu.
Purity is >98%, as determined by Coomassie stained SDS-PAGE.
0.22 µm filtered protein solution is in 10mM NaH2PO4, 150mM NaCl, pH 7.2.
Endotoxin level is <0.1 EU/µg (<0.01ng/µg) protein as determined by the LAL method.
10-100µg sizes are bottled at 200µg/ml. 500µg and larger sizes are bottled at the concentration indicated on the vial.
Storage & Handling:
Unopened vial can be stored at 4°C for three months, at -20°C for six months, or at -70°C for one year. For maximum results, quick spin vial prior to opening. Stock solutions should be prepared at no less than 10µg/mL in buffer containing carrier protein such as 1% BSA or HSA or 10% FBS. For long term storage, aliquot into polypropylene vials and store in a manual defrost freezer. Avoid repeated freeze/thaw cycles.
The ED50 is 0.010-0.020 ng/ml, corresponding to a specific activity of 5-10 X107 units/mg, as determined by a dose dependent stimulation of L929 cells treated with actinomycin D.
Cytotoxic effect on L929 mouse fibroblast cells induced by mouse TNF-α in the presence of actinomycin D.
TNF-α is secreted by macrophages, monocytes, neutrophils, T-cells (principally CD4+), and NK-cells. Many transformed cell lines also secrete TNF-α. TNF-α forms multimeric complexes; stable trimers are most common in solution. A 26 kD membrane form of TNF-α has also been described. TNF-α binding to surface receptors elicits a wide array of biologic activities including: cytolysis and cytostasis of many tumor cell lines in vitro, hemorraghic necrosis of tumors in vivo, increased fibroblast proliferation, and enhanced chemotaxis and phagocytosis in neutrophils.
Type II integral membrane protein processed by TACE for secretion; upregulated by interferons, IL-2, GM-CSF, substance P, bradykinin, PAF, immune complexes, cyclooxygenase; downregulated by IL-6, TGF-β, vitamin D3, prostaglandin E2, PAF antagonists
Activated monocytes, neutrophils, macrophages, T cells, B cells, NK cells, LAK cells
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