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Zinquin ethyl ester *UltraPure grade*
Zinquin ethyl ester is a lipophilic, zinc-sensitive, cell-permeable fluorescent probe. It is retained in living cells because the ethyl ester is cleaved by cytosolic esterase to give Zinquin that carry a negative charge, preventing its efflux across the plasma membrane. Zinquin fluorescent probes may be loaded into cells by in the culture medium containing 5-40 uM Zinquin ethyl ester in PBS with calcium and magnesium (or in culture medium). Cells are normally incubated with the Zinquin ethyl ester for 15-30 minutes at 37 °C. Exact loading concentration, time and temperature depend on the purpose of the experiment and cell type, and so will need to be optimized experimentally. Cells are washed in PBS with culture medium to remove extracellular remaining dye. Cells are observed under microscope or used for confocal microscopy, FACS or spectrofluorimetry analysis. Zinc is the second most abundant transition metal in the body and it is essential as catalytic, structural and regulatory ion. Zinc ions are involved in homeostasis, immune responses, oxidative stress, apoptosis and aging. Zinc has been proposed to function as a conventional neurotransmitter for the presynaptic neuron and as a transmembrane signal to traverse the postsynaptic neuron. Aberrant zinc metabolism is associated with many neurological diseases including Alzheimer's disease, Parkinson's disease and epilepsy. The most suitable technique for in vivo monitoring of zinc has been proven to be fluorescence imaging.
Example protocol
PREPARATION OF STOCK SOLUTION
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.
1. Zinquin stock solution:
Make a ~10 mM Zinquin ethyl ester stock solution with DMSO.
SAMPLE EXPERIMENTAL PROTOCOL
- Load into cells in culture medium containing 5 - 40 uM zinquin ethyl ester or in PBS (with calcium and magnesium).
- Incubate with the zinquin ethyl ester for 15 - 30 minutes at 37°C. Exact loading concentration, time, and temperature depend on the purpose of the experiment and cell type, and so will need to be optimized experimentally.
- Wash with PBS to remove any extracellular remaining dye, and replace with PBS or medium.
- Observe under microscope or analyze with flow cytometer with 515/30 nm filter.
Calculators
Common stock solution preparation
Table 1. Volume of DMSO needed to reconstitute specific mass of Zinquin ethyl ester *UltraPure grade* 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 | 241.272 µL | 1.206 mL | 2.413 mL | 12.064 mL | 24.127 mL |
| 5 mM | 48.254 µL | 241.272 µL | 482.544 µL | 2.413 mL | 4.825 mL |
| 10 mM | 24.127 µL | 120.636 µL | 241.272 µL | 1.206 mL | 2.413 mL |
Molarity calculator
Enter any two values (mass, volume, concentration) to calculate the third.
| Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
| / | = | x | = |
Spectrum
Citations
View all 1 citations: Citation Explorer
Oyster-Derived Zinc-Binding Peptide Modified by Plastein Reaction via Zinc Chelation Promotes the Intestinal Absorption of Zinc
Authors: Li, Jianpeng and Gong, Chen and Wang, Zaiyang and Gao, Ruichang and Ren, Jiaoyan and Zhou, Xiaodong and Wang, Haiyan and Xu, He and Xiao, Feng and Cao, Yuhui and others, undefined
Journal: Marine drugs (2019): 341
Authors: Li, Jianpeng and Gong, Chen and Wang, Zaiyang and Gao, Ruichang and Ren, Jiaoyan and Zhou, Xiaodong and Wang, Haiyan and Xu, He and Xiao, Feng and Cao, Yuhui and others, undefined
Journal: Marine drugs (2019): 341
References
View all 8 references: Citation Explorer
Zinquin identifies subcellular compartmentalization of zinc in cortical neurons. Relation to the trafficking of zinc and the mitochondrial compartment
Authors: Colvin RA, Laskowski M, Fontaine CP.
Journal: Brain Res (2006): 1
Authors: Colvin RA, Laskowski M, Fontaine CP.
Journal: Brain Res (2006): 1
S-Nitroso compounds interfere with zinc probing by Zinquin
Authors: Jansen S, Arning J, Dulcks T, Beyersmann D.
Journal: Anal Biochem (2004): 145
Authors: Jansen S, Arning J, Dulcks T, Beyersmann D.
Journal: Anal Biochem (2004): 145
Coordination and fluorescence of the intracellular Zn2+ probe [2-methyl-8-(4-toluenesulfonamido)-6-quinolyloxy]acetic acid (Zinquin A) in ternary Zn2+ complexes
Authors: Hendrickson KM, Geue JP, Wyness O, Lincoln SF, Ward AD.
Journal: J Am Chem Soc (2003): 3889
Authors: Hendrickson KM, Geue JP, Wyness O, Lincoln SF, Ward AD.
Journal: J Am Chem Soc (2003): 3889
Fluorescent detection of Zn(2+)-rich vesicles with Zinquin: mechanism of action in lipid environments
Authors: Snitsarev V, Budde T, Stricker TP, Cox JM, Krupa DJ, Geng L, Kay AR.
Journal: Biophys J (2001): 1538
Authors: Snitsarev V, Budde T, Stricker TP, Cox JM, Krupa DJ, Geng L, Kay AR.
Journal: Biophys J (2001): 1538
The synthesis and fluorescent properties of analogues of the zinc(II) specific fluorophore zinquin ester
Authors: Kimber MC, Mahadevan IB, Lincoln SF, Ward AD, Tiekink ER.
Journal: J Org Chem (2000): 8204
Authors: Kimber MC, Mahadevan IB, Lincoln SF, Ward AD, Tiekink ER.
Journal: J Org Chem (2000): 8204
Page updated on November 15, 2024
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