Recombinant DNA technology allows the addition of short pieces of well-defined tags, "peptides" or proteins at the N- or C-terminus of target genes, which can provide 'affinity handles' designed to bind specific matrices. Therefore, tags enable a selective identification and purification of the protein of interest. Eukaryotic genes are often cloned into E. coli thioredoxin gene, resulting in the expression of a desired protein as a fusion hybrid with thioredoxin. Thioredoxin is a small electron transport protein that serves as hydrogen door in the enzymatic reduction of ribonucleotides to deoxyribonucleotides. Thioredoxin from E. coli consists of a single polypeptide of 108aa (~11.7kD). The gene encoding Thioredoxin has been clones into several plasmids and serves as a fusion partner for the expression of other clones genes. The system often produces high expression of soluble fusion proteins in the E. coli cytoplasm. The fusion proteins may also fold correctly with biological activity. Anti-Thioredoxin antibodies allow a simple isolation of fusion proteins directly from the crude bacterial lysate, using immunoaffinity chromatography or used in immunoprecipitation. Anti- thioredoxin can also be used for the immunocytochemical detection of thioredoxin in cells and tissues that express transfected bacterial thioredoxin gene. Ant-thioredoxin may be used in various immunoassays to identify the expression of thioredoxin fusion protein.ApplicationsSuitable for use in Western Blot, ELISA and Immunohistochemistry. Other applications not tested. Recommended Dilutions: |Western Blot: 10-50ug/ml|Immunohistochemistry: Frozen and paraffin sections|Optimal dilutions to be determined by the researcher. Recommended Secondary Reagents: |F(ab')2 rabbit anti-mouse IgG (HRP)Storage and StabilityMay be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.Note: Applications are based on unconjugated antibody.ImmunogenRecombinant human thioredoxinFormSupplied as a liquid in PBS, pH 7.2. Labeled with Alkaline Phosphatase.PurityPurifiedSpecificityRecognizes human thioredoxin, a 12kD redox protein which has powerful protein disulphide oxidoreductase activity. Thioredoxin has been shown to be involved in various cellular processes, including activation of the transcription factor NF-KB. Species Crossreactivity: MarmosetReferences1. Perkins, A.V., et al., Placenta 16: 635-642 (1995). 2. Lincoln, D. et al. (2003) The thioredoxin-thioredoxin reductase system: over-expression in human cancer. Anticancer Res. 23: 2425-2434. 3. Stantchev, T.S. et al. (2012) Cell-type specific requirements for thiol/disulfide exchange during HIV-1 entry and infection. Retrovirology. 9: 97. 4. Ma, X. et al. (2001) Regulation of interferon and retinoic acid-induced cell death activation through thioredoxin reductase. J Biol Chem. 276: 24843-54. 5. Ramanathan, R.K. et al. (2007) A Phase I pharmacokinetic and pharmacodynamic study of PX-12, a novel inhibitor of thioredoxin-1, in patients with advanced solid tumors. Clin Cancer Res. 13: 2109-14. 6. Arnold, N.B. et al. (2004) Thioredoxin is downstream of Smad7 in a pathway that promotes growth and suppresses cisplatin-induced apoptosis in pancreatic cancer. Cancer Res. 64: 3599-606.7. Kinoshita, T. et al. (2007) Thioredoxin prevents the development and progression of elastase-induced emphysema. Biochem Biophys Res Commun. 354: 712-9. 8. Dorion, S. et al. (2002) Activation of the p38 signaling pathway by heat shock involves the dissociation of glutathione S-transferase Mu from Ask1.J Biol Chem. 277: 30792-7. 9. Sensi, M. et al. (2005) Immunogenicity without immunoselection: a mutant but functional antioxidant enzyme retained in a human metastatic melanoma and targeted by CD8(+) T cells with a memory phenotype. Cancer Res. 65: 632-40. 10. Mamoon, N.M. et al. (2007) Multiple cysteine residues are implicated in Janus kinase 2-mediated catalysis.Biochemistry. 46: 14810-8.|11. Agathanggelou, A. et al. (2003) Identification of Novel Gene Expression Targets for the Ras Association Domain Family 1 (RASSF1A) Tumor Suppressor Gene in Non-Small Cell Lung Cancer and Neuroblastoma. Cancer Res. 63: 5344-51. 12. Di Trapani, G. et al. (1998) Production and secretion of thioredoxin from transformed human trophoblast cells. Mol Hum Reprod. 4: 369-75.13. Lopata, A. et al. (2001) Expression and localization of thioredoxin during early implantation in the marmoset monkey. Mol Hum Reprod. 7: 1159-65.

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