| Purification and Quality Control | The His-tag recombinant protein is purified by affinity chromatography in combination with FPLC columns. The purified TNF-alpha is greater than 95% homogeneous based on SDS-PAGE analysis. | | Unit Definition (Activity) | 1 unit equals 1 nanogram of purified protein. | | Applications | Recombinant TNFα can be used in cytotoxicity assays and other cell assays. | | Formulation and Storage | The protein is in 20mM Tris-HCl pH7.9,100mM NaCl, 0.2mM EDTA, 1mM DTT and 20% glycerol. Stored at -70°C before use. Avoid repeated freeze thaw cycles. | | Synonym | DIF; TNF-alpha; TNFA and TNFSF2. | | Protein Sequence | VRSSSRTPSD KPVAHVVANP QAEGQLQWLN RRANALLANG VELRDNQLVV PSEGLYLIYS
QVLFKGQGCP STHVLLTHTI SRIAVSYQTK VNLLSAIKSP CQRETPEGAE AKPWYEPIYL
GGVFQLEKGD RLSAEINRPD YLDFAESGQV YFGIIAL | | Background | Tumor necrosis factor alpha (TNFα or TNFSF2) is produced by many different cell types including activated monocytes, fibroblasts, endothelial cells, macrophages, T-cells, B-lymphocytes, granulocytes, smooth muscle cells, eosinophilis, chondrocytes and other cell types (1). It functions as a cytokine involved in cell proliferation, differentiation, apoptosis, lipid metabolism and coagulation (2, 3). High level of TNFα leads to the development of inflammatory responses that are hallmarks of many diseases including autoimmune diseases, insulin resistance and cancer (4, 5). The active form of TNFa (18kDa) is generated by the cleavage of an Ala-Val bond between residues 76-77 of the precursor protein (6). | | References | 1. Locksley, R. et al., (2001) Cell 104, 487-501
2. Mukhopadhyay, S. et al., (2006) Respiratory Res. 7, 1-9
3. Aggarwal, B. (2003) Nat. Rev. Immunol. 3, 745-756
4. Walczak, H. et al., (1999) Nat. Med. 5, 157-163
5. Lam, J. et al., (2000) J. Clin. Invest. 106, 1481-1488
6. Mcgeehan, G. et al., (1994) Nature 370, 558-561 | | Image of SDS-PAGE /Western-blot |  |
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