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OxiSelect Intracellular ROS Assay Kit (Green Fluorescence)

MBS168048

20 Assays

340 USD

Assay Kit

OxiSelect Intracellular ROS Assay Kit (Green Fluorescence)

[OxiSelect Intracellular ROS]

[OxiSelect Intracellular ROS Assay Kit (Green Fluorescence)]

Upon receipt, store the DCFH-DA and DCF Standard at -20 degree C. Avoid multiple freeze/thaw cycles. Store the Cell Lysis Buffer and Hydrogen Peroxide at 4 degree C.

Typical Testing Data/Standard Curve (for reference only)

Oxidative Stress/ Damage; Reactive Oxygen Species (ROS) Assays; Intracellular ROS Assay

. Principle of the assay: The OxiSelect Intracellular ROS Assay Kit is a cell-based assay for measuring antioxidant or ROS activity. Cells are cultured in a 96-well cell culture plate and then pre-incubated with DCFH-DA, which is cell-permeable (Figure 1). The unknown antioxidant or ROS samples are then added to the cells. After a brief incubation, the cells can be read on a standard fluorescence plate reader. The ROS or antioxidant content in unknown samples is determined by comparison with the predetermined DCF standard curve. Background: Accumulation of reactive oxygen species (ROS) coupled with an increase in oxidative stress has been implicated in the pathogenesis of several disease states. The role of oxidative stress in vascular diseases, diabetes, renal ischemia, atherosclerosis, pulmonary pathological states, inflammatory diseases, and cancer has been well established. Free radicals and other reactive species are constantly generated in vivo and cause oxidative damage to biomolecules, a process held in check by the existence of multiple antioxidant and repair systems as well as the replacement of damaged nucleic acids, proteins and lipids. Measuring the effect of antioxidant therapies and ROS activity intracellularly is crucial to suppressing or treating oxidative stress inducers. OxiSelect Intracellular ROS Assay Kit (Green Fluorescence) is a cell-based assay for measuring hydroxyl, peroxyl, or other reactive oxygen species activity within a cell. The assay employs the cell-permeable fluorogenic probe 2', 7'-Dichlorodihydrofluorescin diacetate (DCFH-DA). In brief, DCFH-DA is diffused into cells and is deacetylated by cellular esterases to non-fluorescent 2', 7'-Dichlorodihydrofluorescin (DCFH), which is rapidly oxidized to highly fluorescent 2', 7'-Dichlorodihydrofluorescein (DCF) by ROS (Figure 1). The fluorescence intensity is proportional to the ROS levels within the cell cytosol. The effect of antioxidant or free radical compounds on DCF-DA can be measured against the fluorescence of the provided DCF standard. The kit has a DCF detection sensitivity limit of 10 pM. Each kit provides sufficient reagents to perform up to 96 assays, including standard curve and unknown samples.

1. Bass DA, Parce JW, Dechatelet LR, Szejda P, Seeds MC, Thomas M. Flow cytometric studies of oxidative product formation by neutrophils: A graded response to membrane stimulation. J Immunol. 1983; 130:1910-1917. 2. Brandt R, Keston AS. Synthesis of diacetyldichlorofluorescin: A stable reagent for fluorometric analysis. Anal Biochem. 1965; 11:6-9. 3. Keston AS, Brandt R. The fluorometric analysis of ultramicro quantities of hydrogen peroxide. Anal Biochem.1965; 11:1-5. 1. Warren, G. et al. (2015). Amphiphilic cationic nanogels as brain-targeted carriers for activated nucleoside reverse transcriptase inhibitors. J Neuroimmune Pharmacol. doi:10.1007/s11481-014-9576-7. 2. Fung, S. Y. et al. (2015). Molecular mechanism of cell death induced by King Cobra (Ophiophagus hannah) venom L-amino acid oxidase. Toxicon. 96:38-45. 3. Lolicato, F. et al. (2015). The cumulus cell layer protects the bovine maturing oocyte against fatty acid-induced lipotoxicity. Biol Reprod. doi: 10.1095/biolreprod.114.120634. 4. Planavila, A. et al. (2014). Fibroblast growth factor 21 protects the heart from oxidative stress. Cardiovasc Res. doi: http://dx.doi.org/10.1093/cvr/cvu263. 5. Tummala, K. S. et al. (2014). Inhibition of de novo NAD+ synthesis by oncogenic URI causes liver tumorigenesis through DNA damage. Cancer Cell. 26:826-839. 6. Guo, S. et al. (2014). Control of antioxidative response by the tumor suppressor protein PML through regulating Nrf2 activity. Mol Biol Cell. 25:2485-2498. 7. Li, R. W. et al. (2014). Uptake and protective effects of ergothioneine in human endothelial cells. J Pharmacol Exp Ther. 350:691-700. 8. Peng, S. et al. (2014). In Barrett's esophagus patients and barrett's cell lines, ursodeoxycholic acid increases antioxidant expression and prevents DNA damage by bile acids. Am J Physiol Gastrointest Liver Physiol. 307:G129-G139. 9. Song, H. et al. (2014). Group VIA Phospholipase A2 mitigates palmitate-induced beta-Cell mitochondrial injury and apoptosis. J Biol Chem. 289:14194-14210. 10. Lee, S.O. et al. (2014). The orphan nuclear receptor NR4A1 (Nur77) regulates oxidative and endoplasmic reticulum stress in pancreatic cancer cells. Mol. Cancer Res. 12:527-538. 11. Yang, Y. et al. (2014). IGF-I regulates redox status in breast cancer cells by activating the amino acid transport molecule xC-. Cancer Res. 74:2295-2305. 12. Koontz, J. et al. (2014). Competition through dimerization between antiapoptotic and proapoptotic HS-1-associated protein X-1 (Hax-1). J. Biol. Chem. 289:3468-3477. 13. Kim, E. Y. et al. (2013). NOX2 interacts with podocyte TRPC6 channels and contributes to their activation by diacylglycerol: essential role of podocin in formation of this complex. Am J Physiol Cell Physiol. 305:C960-C971. 14. Abe, Y. et al. (2013). TGF-beta1 stimulates mitochondrial oxidative phosphorylation and generation of reactive oxygen species in cultured mouse podocytes, mediated in part by the mTOR pathway. Am J Physiol Renal Physiol. 305:F1477-F1490. 15. He, Q. et al. (2013). Tafazzin knockdown interrupts cell cycle progression in cultured neonatal ventricular fibroblasts. Am J Physiol Heart Circ Physiol. 305:H1332-H1343. 16. Kokkinaki, M. et al. (2013). Klotho regulates retinal pigment epithelial functions and protects against oxidative stress. J. Neurosci. 33:16346-16359. 17. Hagan, C. et al. (2013). A common docking domain in progesterone receptor-B links DUSP6 and CK2 signaling to proliferative transcriptional programs in breast cancer cells. Nucleic Acids Res. 10.1093/nar/gkt706. 18. Teng, H. et al. (2013). Oxygen-sensitive mitochondrial accumulation of cystathionine beta-synthase mediated by lon protease. PNAS. 110: 12679-12684. 19. Montalvo-Ortiz, B.L. et al. (2012). Characterization of EHop-016, novel small molecule inhibitor of Rac GTPase. J. Biol. Chem. 287:13228-13238. 20. Wang, H. et al. (2012). p53-induced gene 3 mediates cell death induced by glutathione peroxidase 3. J. Biol. Chem. 287:16890-16902. 21. Pierce, L.M. et al. (2012). Effect of heavy metal tungsten alloy particles on oxidative product formation and phagocytosis by lung macrophages. Am. J. Respir. Crit. Care Med. 185:A4666. 22. Druz, A. et al. (2012). Glucose depletion activates mmu-miR-466h-5p expression through oxidative stress and inhibition of histone deacetylation. Nucleic Acids Res. 40:7291-7302. 23. Momi, S. et al. (2012). Nitric oxide enhances the anti-inflammatory and anti-atherogenic activity of atorvastatin in a mouse model of accelerated atherosclerosis. Cardiovasc. Res. 94: 428-438. 24. Cho, K.A. et al. (2012). IL-17 and IL-22 enhance skin inflammation by stimulating the secretion of IL-1beta by keratinocytes via the ROS-NLRP3-Caspase-1 pathway. Int. Immunol. 24:147-158. 25. Martin, S.A. et al. (2011). Parallel high-throughput RNA interference screens identify PINK1 as a potential therapeutic target for the treatment of DNA mismatch repair-deficient cancers. Cancer Res. 71:1836-1848. 26. Kuznetsov, J.N. et al. (2011). AMPK and Akt determine apoptotic cell death following perturbations of one-carbon metabolism by regulating ER Stress in acute lymphoblastic leukemia. Mol. Cancer Ther. 10:437-447. 27. Wei, J. et al. (2011). C-Jun N-terminal Kinase (JNK-1) confers protection against brief but not extended ischemia during acute myocardial infarction. J. Biol. Chem. 286:13995-14006. 28. Titos, E. et al. (2011). Resolvin D1 and its precursor docosahexaenoic acid promote resolution of adipose tissue inflammation by eliciting macrophage polarization toward an M2-like phenotype. J. Immunol. 187:5408-5418. 29. Zhang, J. et al. (2011). 2-Deoxy-D-Glucose attenuates isoflurane-induced cytotoxicity in an in vitro cell culture model of H4 human neuroglioma cells. Anesth. Analg. 113:1468-1475. 30. Zhang, Y. et al. (2010). The mitochondrial pathway of anesthetic isofluorane-induced apoptosis. J. Biol. Chem. 285:4025-4037. 31. Batova, A. et al. (2010). The synthetic caged garcinia xanthone cluvenone induces cell stress and apoptosis and has immune modulatory activity. Mol. Cancer Ther. 9:2869-2878. 32. Tanaka, N. et al. (2010). Cis-dichlorodiammineplatinum upregulates Angiotensin II Type 1 receptors through reactive oxygen species generation and enhances VEGF production in bladder cancer. Mol. Cancer Ther. 9:2982-2992

20 Assays

340 USD

96 Assays

615

5x96 Assays

2430