Chemical Name: Copper(2+), [N-(3',6'-bis(acetyloxy)-3-oxospiro[isobenzofuran-1(3H),9'-[9H]xanthen]-5-yl)-1,4,7,10-tetraazacyclododecane-1-acetamide-κN1,κN4,κN7,κN10]-Coordination Compound
Appearance: Blue solid
Purity: ≧ 90.0 %
Storage Condition: -20℃
Shipping Condition: Ambient temperature
It has been recognized that hydrogen sulfide (H2S) has an important role as a physiological active substance for
vasodilation, cytoprotection, and modulation of insulin secretion. H
2S is considered as a gaseous molecule such as
nitric oxide and carbon monoxide. However, around 80% of the total sulfide exists as hydrogen sulfide anion (HS-
physiological condition, since the pKa is about 7. In addition, HS-
easily converts to various biochemical molecules
such as persulfides and polysulfides, which react with sulfhydryl moieties in a living body. -SulfoBiotics- HSip-1 is a novel
fluorescent probe to detect H
2S selectively and it emits strong green fluorescence when it reacts with H2S. -SulfoBiotics-HSip-1
DA is cell membrane permeable and it enables fluorescent imaging of intracellular H
Fig. 1 Chemical structure of HSip-1 DA
Fig. 2 Excitation and emission spectra of HSip-1 reacted with H2S
(Em: 491nm/Ex: 516nm)
Fluorescence imaging of hydrogen sulfide with HSip-1 DA:
1) HeLa cells were seeded on μ-slide 8 well (Ibidi) and cultured at 37℃ overnight in a 5% CO2 incubator.
2) The culture medium was discarded and the cells were washed with a serum-free medium (MEM) twice.
3) HSip-1 DA stock solution (1 mmol/l) was diluted with a serum-free medium (MEM) to prepare 5 μmol/l HSip-1 DA working solution.
*Please optimize the final concentration of HSip-1 DA depeneding on the cell lines.
4) HSip-1 DA working solution (5 μmol/l, 200 μl) was added to the cells, and the cells were cultured at 37℃ for 30 minutes in a 5% CO2 incubator.
5) The supernatant was discarded, and the cells were washed with HBSS twice.
6) Na2S solution (200 μmol/l, 200 μl) was added to the each well, and the cells were cultured at 37℃ for 30 minutes in a 5% CO2 incubator.
7) The supernatant was discarded and the cells were washed with HBSS twice.
8) HBSS (200 μl) were added, and the cells were observed by confocal fluorescence microscopy
Fig. Detection of hydrogen sulfide using HSip-1 DA in HeLa cells treated with Na2S.
(A: Control, B: 200 μmol/l Na2S treated)
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