DAX-J2™ Ratio 580/460

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Additional ordering information

Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
International	See distributors
Bulk request	Inquire
Custom size	Inquire
Shipping	Standard overnight for United States, inquire for international

Physical properties

Molecular weight	830.97
Solvent	DMSO

Spectral properties

Excitation (nm)	404
Emission (nm)	468

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure
UNSPSC	12352200

Overview SDS Protocol

Molecular weight

830.97

Excitation (nm)

404

Emission (nm)

468

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 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

Molarity calculator

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Spectrum

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Spectral properties

Excitation (nm)	404
Emission (nm)	468

Images

Figure 1. 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).

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
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Journal: Am J Physiol Heart Circ Physiol (2011): H1788

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Journal: Plant Sci (2011): 593

Rapid upregulation of cytoprotective nitric oxide in breast tumor cells subjected to a photodynamic therapy-like oxidative challenge
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Journal: Photochem Photobiol (2011): 378

Sleep deprivation triggers inducible nitric oxide-dependent nitric oxide production in wake-active basal forebrain neurons
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Journal: J Neurosci (2010): 13254

Production and scavenging of nitric oxide by barley root mitochondria
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Journal: Plant Cell Physiol (2010): 576

Production of Nitric Oxide within the Aplysia Californica Nervous System
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Journal: ACS Chem Neurosci (2010): 182