HS-CyBz
HS-CyBz is a Fluorescent probe for H₂S detection, enabling ratiometric optical/photoacoustic dual-modality in/ex vivo imaging. Its detection mechanism relies on nucleophilic substitution of its benzoic ester group by HS⁻, which releases an enolic meso-hydroxyltricarboheptamethine cyanine that then undergoes keto-enol tautomerization to form Cy-ketone; this tautomerization causes distinct shifts in absorption and emission spectra, producing a ratiometric response that reduces interferences from tissue scattering, autofluorescence, and probe concentration. In its initial state, HS-CyBz has an excitation wavelength of 595 nm, with emission bands centered at 805 nm (main) and 630 nm (minor); upon reaction with H₂S, the 805 nm emission band decreases while the 630 nm band drastically increases, and its absorption spectrum shows a sharp band at 775 nm and a shoulder band at 708 nm, which decrease upon H₂S reaction with a minor increase at 850 nm and an isosbestic point at 825 nm. For in vivo optical imaging, excitation at 560 nm is used with emission channels at 620 nm and 790 nm, while in vivo photoacoustic imaging uses excitation at 775 nm and 825 nm. The detection limit of HS-CyBz for H₂S is 0.5 μM, and it shows high selectivity, with only H₂S inducing a distinct enhancement of the emission ratio F₆₃₀/F₈₀₅ and PA ratio PA₈₂₅/PA₇₇₅, while other biochemical species including cations, anions, reactive oxygen species, biothiols, and carboxylesterase trigger only minor changes and do not interfere with H₂S sensing. Tail intravenous injection of HS-CyBz leads to accumulation in the liver of mice, and it can be used to verify endogenous H₂S upregulation triggered by S-adenosyl-L-methionine via ratiometric optical/photoacoustic imaging.
For research use only. We do not sell to patients.
- CAS No.: 2410296-16-9
- Formula: C41H44ClIN2O2
- Molecular Weight:759.16
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Storage:
Please store the product under the recommended conditions in the Certificate of Analysis.
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Biological Activity
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Chemical Information
- Protocols
- Purity & Documentation
- References
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Help & FAQs
Biological Activity
Description
In Vitro
Operating Instructions (The following is a recommended experimental protocol for guidance only, and adjustments are required based on specific requirements)
1. Stock Solution Preparation
2. Working Solution Preparation
2.1 Working concentration: 20 μM.
2.2 Notes: Adjust the working solution concentration as needed; prepare fresh immediately before use.
3. Staining Procedure
3.1 Sample types: Live mouse tissues; ex vivo mouse liver tissues[1].
3.2 Incubation conditions:
3.2.1 For mouse subcutaneous tissue imaging: Use 20 μM probe solution, and incubate for 30 min after analyte injection.
3.2.2 For in vivo/ex vivo mouse liver imaging: Inject the probe intravenously 12 h after intraperitoneal injection of SAM.
3.3 Washing step: No washing step is specified.
4. Control Setup
4.2 Set up negative controls, positive controls and blank controls.
4.2.1 Negative control: Inject normal saline into parallel mouse tissue regions instead of Na2S or SAM.
4.3 Positive controls are used to validate the experimental system; blank controls are used to eliminate fluorescent interference from reagents.
5. Detection and Analysis
5.1 Instruments:
5.1.1 Fluorescence imaging: Ex 560 nm, Em 620 nm and 790 nm; Ex 720 nm, Em 790 nm.
5.1.2 Photoacoustic imaging: Ex 775 nm and 825 nm.
5.2 Result analysis:
5.2.1 Fluorescence intensity changes: In response to H2S, the fluorescence intensity at 620 nm increases while that at 790 nm decreases, leading to an increased F620/F790 ratio.
5.2.2 Photoacoustic signal changes: In response to H2S, the PA signal at 775 nm decreases, leading to an increased PA825/PA775 ratio.
5.2.3 Fluorescence localization: After intravenous injection, the probe accumulates in mouse liver tissues.
5.2.4 Expected ratio changes: SAM induces upregulation of endogenous H2S in mouse liver, resulting in higher F620/F790 and PA825/PA775 ratios compared with the normal saline-treated control group.
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
Chemical Information
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CAS No. 2410296-16-9
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Molecular Weight 759.16
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Formula C41H44ClIN2O2
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SMILES
CCN1C2=C(C(C)(/C1=C\C=C3CCCC(/C=C/C4=[N+](C5=C(C4(C)C)C=CC=C5)CC)=C/3OC(C6=C(C=CC=C6)I)=O)C)C=CC=C2.[Cl-]
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
Please store the product under the recommended conditions in the Certificate of Analysis.
Protocols
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How to Select the Route of Administration for Mammals
Route-of-administration selection in mammals is a pharmacokinetic, pharmacodynamic, formulation, animal-welfare, and translational decision, not a default technical choice. The selected route should match the study goal: intravenous dosing is most useful when complete systemic exposure and rapid onset are required, oral dosing is most translational for orally intended medicines but is affected by absorption and first-pass metabolism, subcutaneous or intramuscular dosing can provide slower systemic exposure, and intraperitoneal dosing can be useful in rodent proof-of-concept studies but may have limited clinical translation. Published route-comparison studies show that the same compound can produce different exposure, onset, bioavailability, tissue distribution, and tolerability depending on route; therefore, route choice should be supported by pilot pharmacokinetic or pharmacodynamic evidence when the literature is insufficient. Unresolved questions include how to standardize route sel
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ROS/oxidative-stress fluorescent staining
ROS/oxidative-stress fluorescent staining uses cell-permeant fluorogenic probes that become fluorescent after oxidation inside cells or tissues; commonly used examples include DCFH-DA/DCFDA for broad cellular oxidant detection, DHE for superoxide-related signal detection, MitoSOX for mitochondrial superoxide-related signal detection, and CellROX probes for oxidative-stress-associated fluorescence readouts. The assay detects probe oxidation rather than a single ROS species unless the probe and analysis method have been chemically validated for that species. DCFH-DA enters cells, is deacetylated by intracellular esterases to DCFH, and produces fluorescent DCF after oxidation, so the readout is used as an operational measure of total cellular oxidative stress rather than a species-specific ROS measurement. DHE and MitoSOX can report superoxide-related oxidation, but red fluorescence alone can include non-specific ethidium-like oxidation products; HPLC or optimized spectral approaches are
Purity & Documentation
References
Calculators
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
HS-CyBz Related Classifications
HY-D3133 Related Classifications
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