Testing Data
OxiSelect In Vitro ROS/RNS Assay Kit
OxiSelect In Vitro ROS/RNS Assay Kit (Green Fluorescence)
Synonyms
OxiSelect In Vitro ROS/RNS; OxiSelect In Vitro ROS/RNS Assay Kit (Green Fluorescence); OxiSelect In Vitro ROS/RNS assay kit
Preparation and Storage
Upon receipt, store the DCF-DiOxyQ and DCF Standard at -20 degree C. Avoid multiple freeze/thaw cycles. Store all other components at 4 degree C.
Testing Data
Testing Data
Testing Data #2
Testing Data #2
Related Product Information for OxiSelect In Vitro ROS/RNS assay kit
Principle of the Assay: The OxiSelect™ In Vitro ROS/RNS Assay Kit is an in vitro assay for measuring total ROS/RNS free radical activity. Unknown ROS or RNS samples or standards are added to the wells with a catalyst that helps accelerate the oxidative reaction. After a brief incubation, the prepared DCFH probe is added to all wells and the oxidation reaction is allowed to proceed (Figure 1). Samples are measured fluorometrically against a hydrogen peroxide or DCF standard. The assay is performed in a 96-well fluorescence plate format that can be read on a standard fluorescence plate reader. The free radical content in unknown samples is determined by comparison with the predetermined DCF or hydrogen peroxide standard curve.
Background/Introduction: Reactive oxygen species (ROS) and reactive nitrogen species (RNS) are well-established molecules responsible for the deleterious effects of oxidative stress. Accumulation of free radicals 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, cancer, as well as ageing 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/RNS activity is crucial to suppressing or treating oxidative stress inducers. The OxiSelect™ In Vitro ROS/RNS Assay Kit is an assay for measuring the total free radical presence of a sample. The assay employs a proprietary quenched fluorogenic probe, dichlorodihydrofluorescin DiOxyQ (DCFH-DiOxyQ), which is a specific ROS/RNS probe that is based on similar chemistry to the popular 2', 7'-dichlorodihydrofluorescein diacetate. The DCFH-DiOxyQ probe is first primed with a quench removal reagent, and subsequently stabilized in the highly reactive DCFH form. In this reactive state, ROS and RNS species can react with DCFH, which is rapidly oxidized to the highly fluorescent 2', 7'-dichlorodihydrofluorescein (DCF) (Figure 1). Fluorescence intensity is proportional to the total ROS/RNS levels within the sample. The DCFH-DiOxyQ probe can react with hydrogen peroxide (H2O2), peroxyl radical (ROO?), nitric oxide (NO), and peroxynitrite anion (ONOO-). These free radical molecules are representative of both ROS and RNS, thus allowing for measurement of the total free radical population within a sample. The OxiSelect™ In Vitro ROS/RNS Assay Kit can also be used to evaluate antioxidant's effect on free radicals. The kit has a detection sensitivity limit of 10 pM for DCF and 40 nM for H2O2. Each Trial Size In Vitro ROS/RNS Assay Kit provides sufficient reagents to perform up to 20 assays, including standard curve and unknown samples.
Background/Introduction: Reactive oxygen species (ROS) and reactive nitrogen species (RNS) are well-established molecules responsible for the deleterious effects of oxidative stress. Accumulation of free radicals 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, cancer, as well as ageing 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/RNS activity is crucial to suppressing or treating oxidative stress inducers. The OxiSelect™ In Vitro ROS/RNS Assay Kit is an assay for measuring the total free radical presence of a sample. The assay employs a proprietary quenched fluorogenic probe, dichlorodihydrofluorescin DiOxyQ (DCFH-DiOxyQ), which is a specific ROS/RNS probe that is based on similar chemistry to the popular 2', 7'-dichlorodihydrofluorescein diacetate. The DCFH-DiOxyQ probe is first primed with a quench removal reagent, and subsequently stabilized in the highly reactive DCFH form. In this reactive state, ROS and RNS species can react with DCFH, which is rapidly oxidized to the highly fluorescent 2', 7'-dichlorodihydrofluorescein (DCF) (Figure 1). Fluorescence intensity is proportional to the total ROS/RNS levels within the sample. The DCFH-DiOxyQ probe can react with hydrogen peroxide (H2O2), peroxyl radical (ROO?), nitric oxide (NO), and peroxynitrite anion (ONOO-). These free radical molecules are representative of both ROS and RNS, thus allowing for measurement of the total free radical population within a sample. The OxiSelect™ In Vitro ROS/RNS Assay Kit can also be used to evaluate antioxidant's effect on free radicals. The kit has a detection sensitivity limit of 10 pM for DCF and 40 nM for H2O2. Each Trial Size In Vitro ROS/RNS Assay Kit provides sufficient reagents to perform up to 20 assays, including standard curve and unknown samples.
Product Categories/Family for OxiSelect In Vitro ROS/RNS assay kit
References
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. Schmidt-Heydt, M. et al. (2015). Oxidative stress induces the biosynthesis of citrinin by penicillium verrucosum at the expense of ochratoxin. Int J Food Microbiol. 192:1-6.
2. Khadir, A. et al. (2015). MAP kinase phosphatase DUSP1 is overexpressed in obese humans and modulated by physical exercise. Am J Physiol Endocrinol Metab. 308:E71-E83.
3. Ravassa, S. et al. (2015). Association of low GLP-1 with oxidative stress is related to cardiac disease and outcome in patients with type 2 diabetes mellitus: a pilot study. Free Radic Biol Med. doi: 10.1016/j.freeradbiomed.2015.01.002.
4. Canivet, L. et al. (2015). Effects of engineered iron nanoparticles on the bryophyte, Physcomitrella patens (Hedw.) Bruch & Schimp, after foliar exposure. Ecotoxicol Environ Saf. 113:499-505.
5. Meenalochani, S. et al. (2015). Sphingosine kinase 2 and sphingosine-1-phosphate promotes mitochondrial function in dopaminergic neurons of mouse model of Parkinson's Disease and in MPP+-treated MN9D cells in vitro. Neuroscience. doi: 10.1016/j.neuroscience.2015.01.032.
6. Walsh, C. J. et al. (2015). Sublethal red tide toxin exposure in free-ranging manatees (Trichechus manatus) affects the immune system through reduced lymphocyte proliferation responses, inflammation, and oxidative stress. Aquatic Toxicology. 161:73-84.
7. Rhyu, H. S. et al. (2014). The effects of ketogenic diet on oxidative stress and antioxidative capacity markers of Taekwondo athletes. J Exerc Rehabil. 10:362-366.
8. Hao, Y. et al. (2014). Mycoplasma pneumoniae modulates STAT3-STAT6/EGFR-FOXA2 signaling to induce overexpression of airway mucins. Infect Immun. 82:5246-5255.
9. Pandey, D. et al. (2014). Transcriptional regulation of endothelial arginase 2 by histone deacetylase 2. Arterioscler Thromb Vasc Biol. 34:1556-1566.
10. Liu, X. et al. (2013). Epoxyeicosatrienoic acids prevent cisplatin-induced renal apoptosis through a p38 mitogen-activated protein kinase-regulated mitochondrial pathway. Mol. Pharmacol. 84:925-934.
11. Song, J. et al. (2013). Nicotinamide phosphoribosyltransferase is required for the calorie restriction-mediated improvements in oxidative stress, mitochondrial biogenesis, and metabolic adaptation. J. Gerontol A Biol Sci Med Sci. 10.1093/gerona/glt122.
12. Xiao, D. et al. (2013). Estrogen normalizes perinatal nicotine-induced hypertensive responses in adult female rat offspring. Hypertension. 61:1246-1254.
13. Wang, W. et al. (2012). Mono-(2-ethylhexyl) phthalate induces oxidative stress and inhibits growth of mouse ovarian antral follicles. Biol Reprod. 87:152.
14. Ju, D.J. et al. (2012). Ethyl pyruvate ameliorates albuminuria and glomerular injury in the animal model of diabetic nephropathy. Am J Physiol Renal Physiol. 302: F606-F613.
15. 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. 10.1093/cvr/cvs100.
16. Patterson, A.J. et al. (2011). Hypoxia-derived oxidative stress mediates epigenetic repression of pkc? gene in foetal rat hearts. Cardiovasc Res. 10.1093/cvr/cvr322.
17. Rathnasamy, G. et al. (2011). Iron and iron regulatory proteins in amoeboid microglial cells are linked to oligodendrocyte death in hypoxic neonatal rat periventricular white matter through production of proinflammatory cytokines and reactive oxygen/nitrogen species. J. Neurosci. 31:17982-17995.
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. Schmidt-Heydt, M. et al. (2015). Oxidative stress induces the biosynthesis of citrinin by penicillium verrucosum at the expense of ochratoxin. Int J Food Microbiol. 192:1-6.
2. Khadir, A. et al. (2015). MAP kinase phosphatase DUSP1 is overexpressed in obese humans and modulated by physical exercise. Am J Physiol Endocrinol Metab. 308:E71-E83.
3. Ravassa, S. et al. (2015). Association of low GLP-1 with oxidative stress is related to cardiac disease and outcome in patients with type 2 diabetes mellitus: a pilot study. Free Radic Biol Med. doi: 10.1016/j.freeradbiomed.2015.01.002.
4. Canivet, L. et al. (2015). Effects of engineered iron nanoparticles on the bryophyte, Physcomitrella patens (Hedw.) Bruch & Schimp, after foliar exposure. Ecotoxicol Environ Saf. 113:499-505.
5. Meenalochani, S. et al. (2015). Sphingosine kinase 2 and sphingosine-1-phosphate promotes mitochondrial function in dopaminergic neurons of mouse model of Parkinson's Disease and in MPP+-treated MN9D cells in vitro. Neuroscience. doi: 10.1016/j.neuroscience.2015.01.032.
6. Walsh, C. J. et al. (2015). Sublethal red tide toxin exposure in free-ranging manatees (Trichechus manatus) affects the immune system through reduced lymphocyte proliferation responses, inflammation, and oxidative stress. Aquatic Toxicology. 161:73-84.
7. Rhyu, H. S. et al. (2014). The effects of ketogenic diet on oxidative stress and antioxidative capacity markers of Taekwondo athletes. J Exerc Rehabil. 10:362-366.
8. Hao, Y. et al. (2014). Mycoplasma pneumoniae modulates STAT3-STAT6/EGFR-FOXA2 signaling to induce overexpression of airway mucins. Infect Immun. 82:5246-5255.
9. Pandey, D. et al. (2014). Transcriptional regulation of endothelial arginase 2 by histone deacetylase 2. Arterioscler Thromb Vasc Biol. 34:1556-1566.
10. Liu, X. et al. (2013). Epoxyeicosatrienoic acids prevent cisplatin-induced renal apoptosis through a p38 mitogen-activated protein kinase-regulated mitochondrial pathway. Mol. Pharmacol. 84:925-934.
11. Song, J. et al. (2013). Nicotinamide phosphoribosyltransferase is required for the calorie restriction-mediated improvements in oxidative stress, mitochondrial biogenesis, and metabolic adaptation. J. Gerontol A Biol Sci Med Sci. 10.1093/gerona/glt122.
12. Xiao, D. et al. (2013). Estrogen normalizes perinatal nicotine-induced hypertensive responses in adult female rat offspring. Hypertension. 61:1246-1254.
13. Wang, W. et al. (2012). Mono-(2-ethylhexyl) phthalate induces oxidative stress and inhibits growth of mouse ovarian antral follicles. Biol Reprod. 87:152.
14. Ju, D.J. et al. (2012). Ethyl pyruvate ameliorates albuminuria and glomerular injury in the animal model of diabetic nephropathy. Am J Physiol Renal Physiol. 302: F606-F613.
15. 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. 10.1093/cvr/cvs100.
16. Patterson, A.J. et al. (2011). Hypoxia-derived oxidative stress mediates epigenetic repression of pkc? gene in foetal rat hearts. Cardiovasc Res. 10.1093/cvr/cvr322.
17. Rathnasamy, G. et al. (2011). Iron and iron regulatory proteins in amoeboid microglial cells are linked to oligodendrocyte death in hypoxic neonatal rat periventricular white matter through production of proinflammatory cytokines and reactive oxygen/nitrogen species. J. Neurosci. 31:17982-17995.
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Product Notes
The OxiSelect In Vitro ROS/RNS (Catalog #AAA168257) is an Assay Kit and is intended for research purposes only. The product is available for immediate purchase. It is sometimes possible for the material contained within the vial of "OxiSelect In Vitro ROS/RNS, Assay Kit" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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