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DAX-J2™ Ratio 580/460
DAX-J2™ Ratio 580/460 is a new nitric oxide (NO) sensor recently developed by AAT Bioquest. It is a cell permeable reagent that can measure free NO and nitric oxide synthase (NOS) activity in living cells in a ratiometric mode under physiological conditions. Once inside the cell the blocking groups on the DAX-J2 reagent are released to induce fluorescence ratio changes at wavelengths of 580 and 460 nm upon NO oxidation. The fluorescence intensities at 580 nm and 460 nm can be detected using the filter sets of Cy3®/TRITC and BD Horizon™ V450/Pacific Blue. These two filter sets are equipped with most of flow cytometers and fluorescence microscope. DAX-J2 Ratio 580/460 has distinct advantages for NO detection than the popular DAF-2 NO probe: 1). It does not require esterase activity for NO detection. DAF-2 requires intracellular esterases to cleave its acetate groups for detecting NO activity. This esterase dependence often complicates the NO detection since esterse activities are affected by cell health and many other factors. 2). DAX-J2 product exhibits pH-independent fluorescence while DAF-2 has its fluorescence highly affected by pH. 3). DAX-J2 Ratio 580/460 can be monitored in a ratiometric mode.
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Common stock solution preparation
Table 1. Volume of DMSO needed to reconstitute specific mass of DAX-J2™ Ratio 580/460 to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.
| 0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
| 1 mM | 120.341 µL | 601.706 µL | 1.203 mL | 6.017 mL | 12.034 mL |
| 5 mM | 24.068 µL | 120.341 µL | 240.683 µL | 1.203 mL | 2.407 mL |
| 10 mM | 12.034 µL | 60.171 µL | 120.341 µL | 601.706 µL | 1.203 mL |
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Enter any two values (mass, volume, concentration) to calculate the third.
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Citations
View all 2 citations: Citation Explorer
Fluorescent real-time quantitative measurements of intracellular peroxynitrite generation and inhibition
Authors: Luo, Zhen and Zhao, Qin and Liu, Jixiang and Liao, Jinfang and Peng, Ruogu and Xi, Yunting and Diwu, Zhenjun
Journal: Analytical biochemistry (2017): 44--48
Authors: Luo, Zhen and Zhao, Qin and Liu, Jixiang and Liao, Jinfang and Peng, Ruogu and Xi, Yunting and Diwu, Zhenjun
Journal: Analytical biochemistry (2017): 44--48
Inducible Nitric Oxide Synthase (iNOS) Is a Novel Negative Regulator of Hematopoietic Stem/Progenitor Cell Trafficking
Authors: Adamiak, Mateusz and Abdelbaset-Ismail, Ahmed and Moore, Joseph B and Zhao, J and Abdel-Latif, Ahmed and Wysoczynski, Marcin and Ratajczak, Mariusz Z
Journal: Stem Cell Reviews and Reports (2016): 1--12
Authors: Adamiak, Mateusz and Abdelbaset-Ismail, Ahmed and Moore, Joseph B and Zhao, J and Abdel-Latif, Ahmed and Wysoczynski, Marcin and Ratajczak, Mariusz Z
Journal: Stem Cell Reviews and Reports (2016): 1--12
References
View all 139 references: Citation Explorer
Pitfalls and limitations in using 4,5-diaminofluorescein for evaluating the influence of polyphenols on nitric oxide release from endothelial cells
Authors: Uhlenhut K, Hogger P.
Journal: Free Radic Biol Med (2012): 2266
Authors: Uhlenhut K, Hogger P.
Journal: Free Radic Biol Med (2012): 2266
Effects of moderate electrical stimulation on reactive species production by primary rat skeletal muscle cells: cross talk between superoxide and nitric oxide production
Authors: Lambertucci RH, Silveira Ldos R, Hirabara SM, Curi R, Sweeney G, Pithon-Curi TC.
Journal: J Cell Physiol (2012): 2511
Authors: Lambertucci RH, Silveira Ldos R, Hirabara SM, Curi R, Sweeney G, Pithon-Curi TC.
Journal: J Cell Physiol (2012): 2511
Improved measurements of intracellular nitric oxide in intact microvessels using 4,5-diaminofluorescein diacetate
Authors: Zhou X, He P.
Journal: Am J Physiol Heart Circ Physiol (2011): H108
Authors: Zhou X, He P.
Journal: Am J Physiol Heart Circ Physiol (2011): H108
Aging negatively affects estrogens-mediated effects on nitric oxide bioavailability by shifting ERalpha/ERbeta balance in female mice
Authors: Novensa L, Novella S, Medina P, Segarra G, Castillo N, Heras M, Hermenegildo C, Dantas AP.
Journal: PLoS One (2011): e25335
Authors: Novensa L, Novella S, Medina P, Segarra G, Castillo N, Heras M, Hermenegildo C, Dantas AP.
Journal: PLoS One (2011): e25335
Temporal and spatial correlation of platelet-activating factor-induced increases in endothelial [Ca(2)(+)]i, nitric oxide, and gap formation in intact venules
Authors: Zhou X, He P.
Journal: Am J Physiol Heart Circ Physiol (2011): H1788
Authors: Zhou X, He P.
Journal: Am J Physiol Heart Circ Physiol (2011): H1788
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