Quantitative Assay, suitable for detection of Prx I, II, III, IV, V via Spectrophotometric UV/Vis at 340nm, sufficient for 100 reactions, ultra accurate, ready-to-use, quick protocol, hyper sensitive, high reproducibility
Thioredoxin is a small (12 kDa) ubiquitous protein present in prokaryotes as well as eukaryotes. Thioredoxin maintains the redox pool of the cell, and is known to be induced in conditions of oxidative stress. Thioredoxin is also an anti-oxidative protein that has been shown to provide protection against oxidative stress in cardiovascular diseases, neurodegenerative conditions, and other inflammatory diseases. Increased expression of thioredoxin is known to be associated with increased proliferation of cancer cells. The activity of the thioredoxin system has been reported to be disrupted in various physiological disorders such as HIV infection, Alzheimer disease and neoplastic disease. Therefore, thioredoxin activity is a very useful marker for several diseases. The assay method in the kit is based on the reduction of insulin disulfides by reduced thioredoxin with thioredoxin reductase and NADPH as ultimate electron donor. This assay is simple to use and is optimized to provide a linear rate of reaction.
Thioredoxin is a disulfide reductase which chemically reacts with disulfides converting it to a thiol using NADPH as the source of reducing power. In this process thioredoxin is auto-oxidized. Thioredoxin reductase regenerates the active thiol group of thioredoxin using reducing equivalents from NADPH. The principle of our thioredoxin assay utilizes insulin disulfide as the target for reduction by Trx. One mole of NADPH is required to reduce one mole of insulin disulfide. Therefore, in the assay the rate of disappearance of NADPH is spectrophotometrically measured at 340 nm as a negative slope. The rate of change of absorbance per minute is converted to μmol of NADPH required to reduce insulin disulfide.
The Thioredoxin Activity Assay Kit by Redoxica has undergone extensive testing with a multitude of lysate types, has been published in dozens of leading scientific journals and after a decade since its market debut, remains the undisputed leader in the detection and measurement of Trx activity. We designed the assay by utilizing the core principles of biochemistry to ensure maximum accuracy, sensitivity, and reproducibility accompanied by a quick and easy to use protocol.
|Unit of Measure||Assays|
|Shelf Life||24 MONTHS|
|Made In||United States of America|
|Components||• Thioredoxin Activity Assay Buffer (2 x 50 mL bottles, Store at 2-8°C)
• Insulin (5 mL vial, Store at 2-8°C)
• β-NADPH (2 vials, Store at: -20°C, Use within 4-5 hours of reconstitution, Keep reconstituted NADPH on ice)
• Thioredoxin (Trx) Positive Control (40 μl tube, Store at -20°C)
• Thioredoxin Reductase (TrxR) (300 μl vial; Store at -20°C)
• Activation Buffer (250 μl tube; Store at -20°C)
|Required Materials Not Included||• Spectrophotometer equipped to perform enzyme kinetics with UV/VIS range.
• Quartz micro cuvettes capable of reading 50-300 μl volume in a 1-cm light path.
|Synonyms||Thioredoxin Activity Assay Kit, TRX, TXN, TRDX, TRX1, Surface-Associated Sulphydryl Protein, SASP, ATL-Derived Factor, ADF, TXN Delta, Thioredoxin Delta, RP11-427L11.1, MGC61975, DKFZp686B1993|
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Q. How do I get a lot specific certificate of analysis? A. The latest batch COA is available on the product page of our website. For a prior lot, please submit a request by visiting our "Contact Us" page. Be sure to provide the product code and batch number.
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Cui, L., Wang, Z., Miao, J., Miao, M., Chandra, R., Jiang, H., … Cui, L. (2012). Mechanisms of in vitro resistance to dihydroartemisinin in Plasmodium falciparum. Molecular Microbiology, 86(1), 111–128. http://doi.org/10.1111/j.1365-2958.2012.08180.xview abstract - Pmid: 22812578
Zieger, M. A., Gupta, M. P., & Wang, M. (2011). Proteomic analysis of endothelial cold-adaptation. BMC Genomics, 12, 630. http://doi.org/10.1186/1471-2164-12-630view abstract - Pmid: 22192797
Huang, Q., Zhou, H. J., Zhang, H., Huang, Y., Hinojosa-Kirschenbaum, F., Fan, P., … Min, W. (2015). Thioredoxin-2 Inhibits Mitochondrial ROS Generation and ASK1 Activity to Maintain Cardiac Function. Circulation, 131(12), 1082–1097. http://doi.org/10.1161/CIRCULATIONAHA.114.012725view abstract - Pmid: 25628390
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STORAGE & SAFETY
Recommended to store insulin at 2-8°C (KEEP REFRIGERATED) and all other kit components at - 20°C (KEEP FROZEN, DRY). For lyophilized kit component storage upon reconsistution please refer to the product insert for more information. Avoid repeated freeze-t
Precautions & Disclaimer
For Laboratory Research and Development Use Only. Not for drug, household or other uses.
SHIPPING & HANDLING
Each kit utilizes a variety of packaging materials including borosilicate glass, PET, polybags, magnetic board, crosslinked foam, PVC.
12"" X 12"" X 10""