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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.

Calculators

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 mg0.5 mg1 mg5 mg10 mg
1 mM120.341 µL601.706 µL1.203 mL6.017 mL12.034 mL
5 mM24.068 µL120.341 µL240.683 µL1.203 mL2.407 mL
10 mM12.034 µL60.171 µL120.341 µL601.706 µL1.203 mL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

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Spectrum

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
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

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
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
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
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
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
Page updated on November 20, 2024

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Catalog Number16310
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Physical properties

Molecular weight

830.97

Solvent

DMSO

Spectral properties

Excitation (nm)

404

Emission (nm)

468

Storage, safety and handling

H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22

Storage

Freeze (< -15 °C); Minimize light exposure
UNSPSC12352200
Fluorescence response of DAX-J2TM Ratio 580/460 (2 uM) to different reactive oxygen species (1 mM) in PBS buffer (pH 7.2). The fluorescence intensities were measured at 580nm and 460 nm (Ex=404nm).
Fluorescence response of DAX-J2TM Ratio 580/460 (2 uM) to different reactive oxygen species (1 mM) in PBS buffer (pH 7.2). The fluorescence intensities were measured at 580nm and 460 nm (Ex=404nm).
Fluorescence response of DAX-J2TM Ratio 580/460 (2 uM) to different reactive oxygen species (1 mM) in PBS buffer (pH 7.2). The fluorescence intensities were measured at 580nm and 460 nm (Ex=404nm).