WSP-5 (Synonyms: Washington State Probe-5) |
| رقم الكتالوجGC45162 |
WSP-5 (Washington State Probe-5) is a fluorescent probe containing active disulfide groups, specifically designed for rapid detection of hydrogen sulfide (H2S) in biological samples and within cells.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 1593024-78-2
Sample solution is provided at 25 µL, 10mM.
WSP-5 (Washington State Probe-5) is a fluorescent probe containing active disulfide groups, specifically designed for rapid detection of hydrogen sulfide (H2S) in biological samples and within cells. H2S, like nitric oxide, has significant effects on mammalian immune, nervous, cardiovascular, and pulmonary systems. Through a reaction-based fluorescence turn-on strategy, WSP-5 selectively and rapidly reacts with H2S, generating fluorescence with excitation and emission maxima at 502 and 525 nm, respectively[1]. Compared with the three H2S probes WSP-1, CAY and P3, WSP-5 has the widest linear range of sensitivity, minimal interference to thiols and other analytes, the most stable in different pH environments, and can be used in Imaging H2S in live cell systems [2].
References:
[1] Peng, B., Chen, W., Liu, C., et al. Fluorescent probes based on nucleophilic substitution-cyclization for hydrogen sulfide detection and bioimaging[J]. Chemistry 20(4), 1010-1016 (2014).
[2] Yingzhu Zhou, Dr. Federico Mazur,et al. Sensitivity and Selectivity Analysis of Fluorescent Probes for Hydrogen Sulfide Detection[J]. Chemistry – An Asian Journal
March 1, 2022.
This plan only provides a guide, please modify it to meet your specific needs.
1.Solution Preparation:
(1) Stock Solution Preparation: Equilibrate solid WSP-5 to room temperature and dissolve it in DMSO to prepare a stock solution with a concentration of 10 mM. Note: Unused stock solution should be aliquoted and stored in the dark at -20°C or -80°C to avoid repeated freeze-thaw cycles.
(2) Working Solution Preparation: Dilute the stock solution with an appropriate buffer (e.g., serum-free culture medium or PBS) to the desired working concentration, such as 10-100 μM. Note: Optimal working concentrations should be adjusted based on experimental conditions or consulted literature for gradient concentration settings. Working solutions must be freshly prepared.
2.Detection of Cellular H2S Using WSP-5 [1] (Reference Only)
(1) Cell Treatment: Culture 2.5×10^5 4T1 cancer cells in a 24-well plate and allow them to adhere for 12 hours.
(2) WSP-5 Incubation: Incubate the cells with 100 μM WPS-5 probe for 30 minutes.
(3) Na2S Treatment:Treat the cells with 200 μM Na2S.
(4) Fluorescence Detection:After 10 minutes, acquire fluorescence images using confocal laser scanning microscopy (CLSM). λ ex/em = 502/525 nm.
3.Analysis of Obtained Fluorescence Intensity Data to Evaluate WSP-5 Sensitivity to H2S [2] (Reference Only)
(1) Experimental Preparation: Use black, clear-bottom 96-well plates for the experiment. Prepare a final volume of 200 μL in each well.
Prepare PBS containing 1 mM CTAB as the buffer for all experiments.
(2) Probe Solubilization: Dissolve the WSP-5 probe in PBS containing 1 mM CTAB to a final concentration of 10 μM.
(3) H2S Donor Addition: Add increasing concentrations of Na2S or DAT as the H2S donor to the wells containing the WSP-5 probe. Concentrations range from 0 to 10,000 μM.
(4) Fluorescence Measurement: Measure the fluorescence intensity at regular intervals (e.g., every 30 seconds) using a SpectraMax M5 microplate reader. λex/em = 502/525 nm.
(5) Data Collection: Record the fluorescence intensity measurements over a period of 30 minutes.
Precautions:This experimental protocol provides guidance for detecting cellular H2S using WSP-5 and subsequent fluorescence measurements. Adjustments can be made according to specific experimental requirements.
References:
[1] He T , Qin X , Jiang C ,et al.Tumor pH-responsive metastable-phase manganese sulfide nanotheranostics for traceable hydrogen sulfide gas therapy primed chemodynamic therapy[J].Theranostics, 2020, 10(6):2453-2462.
[2] Yingzhu Zhou, Dr. Federico Mazur,et al. Sensitivity and Selectivity Analysis of Fluorescent Probes for Hydrogen Sulfide Detection[J]. Chemistry – An Asian Journal
March 1, 2022.
| Cas No. | 1593024-78-2 | SDF | |
| المرادفات | Washington State Probe-5 | ||
| Chemical Name | 3-oxo-3H-spiro[isobenzofuran-1,9'-xanthene]-3',6'-diyl bis(2-(pyridin-2-yldisulfanyl)benzoate) | ||
| Canonical SMILES | O=C(OC1=CC=C2C(OC3=C(C24C5=CC=CC=C5C(O4)=O)C=CC(OC(C6=CC=CC=C6SSC7=CC=CC=N7)=O)=C3)=C1)C8=C(SSC9=NC=CC=C9)C=CC=C8 | ||
| Formula | C44H26N2O7S4 | M.Wt | 822.9 |
| الذوبان | 25 mg/ml in DMSO, 30 mg/ml in DMF | Storage | Store at -20°C; protect from light |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
|
1 mg | 5 mg | 10 mg |
| 1 mM | 1.2152 mL | 6.0761 mL | 12.1521 mL |
| 5 mM | 0.243 mL | 1.2152 mL | 2.4304 mL |
| 10 mM | 0.1215 mL | 0.6076 mL | 1.2152 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 15 reference(s) in Google Scholar.)
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