Semi-quantitative, gel based assay, sufficient for 40 reactions, highly efficacious, easy-to-use, reliable, reproducible results.
Peroxiredoxins (Prx) are a family of peroxidase proteins that reduce peroxides in the cells. In addition to reduction of peroxides these proteins are also implicated in many cellular functions such as differentiation, proliferation, and apoptosis. There are six different members of the mammalian peroxiredoxin family which are grouped as 2-Cys or 1-Cys peroxiredoxins. Peroxiredoxin isoenzymes I-V depend on reduced Thioredoxin for their reducing action. These are known as 2-Cys peroxiredoxins. Prx I is known to be involved in tumor suppression as an inhibitor of tyrosine kinase, and has been shown to be overexpressed in lung cancer. The overexpression of Prx I, II, V and VI has been shown to be increased in breast cancer. The levels of Prx I and Prx II has been shown to be significantly increased in Alzheimer’s disease. Recent studies have shown that mammalian peroxiredoxins are inactivated by oxidation of active site cysteine residues. In addition, recent studies have indicated that this oxidation process is reversible. Therefore, determination of activity of Prx is crucial in attributing biological function of Prx in conditions of oxidative stress in cancer, neurodegenerative diseases and in other diseases.
Mammalian 2-Cys peroxiredoxins are cellular peroxidases that reduce peroxides in the presence of Thioredoxin. Thioredoxin is oxidized in the process and is regenerated by Thioredoxin reductase that utilizes reducing equivalents from NADPH. In our assay method E.coli Trx and mammalian Thioredoxin reductase (TrxR) are used to provide reducing power to 2-Cys-Peroxiredoxins for reduction of peroxides such as hydrogen peroxide. The utilization of NADPH for the reduction of peroxides is monitored as the decrease in absorbance of NADPH at 340 nm. Specific activity is the amount of NADPH oxidized per minute per milligram of protein. A reaction blank is utilized in the absence of Thioredoxin to determine the non-specific decrease in the level of NADPH by mammalian TrxR.
Redoxica's 2-Cys-Peroxiredoxin Activity Assay Kit 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 has been established as the undisputed leader in the detection and activity measurement of 2-cys-peroxiredoxin. We developed the assay by utilizing core principles of biochemistry to ensure optimal 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||• Peroxiredoxin Assay Buffer (2 x 50 mL bottles, Store at 2-8°C)
• Thioredoxin (2 x 250 μL vial, Store at -20°C)
• Thioredoxin Reductase (2 x 150 μL vial, Store at -20°C)
• β-NADPH (2 vials, Store at: -20°C, use within 3-4 hours of reconstitution, keep reconstituted NADPH on ice, not recommended for storage)
• Substrate Solution (1mL vial, Store at 2-8°C, light sensitive)
|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. Make sure that you use the right kind of micro-quartz cuvette for a specific spectrophotometer as they may vary.
|Synonyms||Peroxiredoxin Activity Assay Kit, 2-Cys-Peroxiredoxin Activity Assay Kit, Peroxiredoxin 1 Assay Kit, Peroxiredoxin I Assay Kit, Prx 1 Assay Kit, Prx I Assay Kit, Peroxiredoxin 2 Assay Kit, Peroxiredoxin II Assay Kit, Prx 2 Assay Kit, Prx II Assay Kit, Peroxiredoxin 3 Assay Kit, Peroxiredoxin III Assay Kit, Prx 3 Assay Kit, Prx III Assay Kit, Peroxiredoxin 4 Assay Kit, Peroxiredoxin IV Assay Kit, Prx 4 Assay Kit, Prx IV Assay Kit, Peroxiredoxin 5 Assay Kit, Peroxiredoxin V Assay Kit, Prx 5 Assay Kit, Prx V Assay Kit, Peroxiredoxin Assay Kit, Prx Assay Kit, PRDX1 Assay Kit, PRDX2 Assay Kit, PRDX3 Assay Kit, PRDX4 Assay Kit, PRDX5 Assay Kit, PRDX Assay Kit, 2-Cys Prx Assay Kit, 2-Cys PRDX Assay Kit|
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Journal of Neuroscience
Hu, X., Weng, Z., Chu, C. T., Zhang, L., Cao, G., Gao, Y., … Chen, J. (2011). Peroxiredoxin-2 protects against 6-hydroxydopamine-induced dopaminergic neurodegeneration via attenuation of the ASK1 signaling cascade. The Journal of Neuroscience : The Official Journal of the Society for Neuroscience, 31(1), 247–261. http://doi.org/10.1523/JNEUROSCI.4589-10.2011view abstract - Pmid: 21209210
Fiskus, W., Coothankandaswamy, V., Chen, J., Ma, H., Ha, K., Saenz, D. T., … Bhalla, K. N. (2016). SIRT2 deacetylates and inhibits the peroxidase activity of peroxiredoxin-1 to sensitize breast cancer cells to oxidant stress inducing agents. Cancer Research, 76(18), 5467–5478. http://doi.org/10.1158/0008-5472.CAN-16-0126view abstract - Pmid: 27503926
Kim, T. H., Song, J., Kim, S.-H., Parikh, A. K., Mo, X., Palanichamy, K., … Kwon, C.-H. (2014). Piperlongumine treatment inactivates peroxiredoxin 4, exacerbates endoplasmic reticulum stress, and preferentially kills high-grade glioma cells. Neuro-Oncology, 16(10), 1354–1364. http://doi.org/10.1093/neuonc/nou088view abstract - Pmid: 24879047
Biotechnology and Bioengineering
Chow‐shi‐yée, M. , Grondin, M. , Averill‐Bates, D. A. and Ouellet, F. (2016), Plant protein 2‐Cys peroxiredoxin TaBAS1 alleviates oxidative and nitrosative stresses incurred during cryopreservation of mammalian cells. Biotechnol. Bioeng., 113: 1511-1521. doi:10.1002/bit.25921view abstract - Pmid: 26724792
Biochimica et Biophysica Acta
Myung-Chul Chung, Farhang Alem, Sarah G. Hamer, Aarthi Narayanan, Konstantin Shatalin, Charles Bailey, Evgeny Nudler, Ramin M. Hakami, S-nitrosylation of peroxiredoxin 1 contributes to viability of lung epithelial cells during Bacillus anthracis infection, Biochimica et Biophysica Acta (BBA), Volume 1861, Issue 1, Part A, 2017, Pages 3019-3029, ISSN 0304-4165, https://doi.org/10.1016/j.bbagen.2016.09.006. (http://www.sciencedirect.com/science/article/pii/S0304416516303282)view abstract - Pmid: 27612662
Journal of Cellular Biochemistry
Lin, H. D., Fong, C. , Biswas, A. , Choolani, M. and Bongso, A. (2016), Human Umbilical Cord Wharton's Jelly Stem Cell Conditioned Medium Induces Tumoricidal Effects on Lymphoma Cells Through Hydrogen Peroxide Mediation. J. Cell. Biochem., 117: 2045-2055. doi:10.1002/jcb.25501view abstract - Pmid: 27392313
Chu, K. L., Lew, Q. J., Rajasegaran, V., Kung, J. T., Zheng, L., Yang, Q., … Chao, S.-H. (2013). Regulation of PRDX1 peroxidase activity by Pin1. Cell Cycle, 12(6), 944–952. http://doi.org/10.4161/cc.23916view abstract - Pmid: 23421996
Antioxidants & Redox Signaling
Gan, Y., Ji, X., Hu, X., Luo, Y., Zhang, L., Li, P., … Chen, J. (2012). Transgenic Overexpression of Peroxiredoxin-2 Attenuates Ischemic Neuronal Injury Via Suppression of a Redox-Sensitive Pro-Death Signaling Pathway. Antioxidants & Redox Signaling, 17(5), 719–732. http://doi.org/10.1089/ars.2011.4298view abstract - Pmid: 22356734
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STORAGE & SAFETY
Recommended to store assay buffer and substrate solution (LIGHT SENSITIVE) 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
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""