WSP-1 is a selective, fast-responsive H2S-specific fluorescent dye (Ex/Em=465/515 nm). WSP-1 reacts with H2S and releases the fluorophore.

Physicochemical Properties

Molecular Formula	H2PSW
Molecular Weight	591.653
Exact Mass	591.081
CAS #	1352750-34-5
PubChem CID	101544503
Appearance	White to off-white solid powder
Density	1.5±0.1 g/cm3
Boiling Point	769.9±60.0 °C at 760 mmHg
Flash Point	419.4±32.9 °C
Vapour Pressure	0.0±2.6 mmHg at 25°C
Index of Refraction	1.762
LogP	6.14
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	9
Rotatable Bond Count	7
Heavy Atom Count	42
Complexity	984
Defined Atom Stereocenter Count	0
InChi Key	VHKQAUGSGQUCDU-UHFFFAOYSA-N
InChi Code	InChI=1S/C33H21NO6S2/c1-37-20-13-15-25-27(18-20)39-28-19-21(14-16-26(28)33(25)24-10-4-2-8-22(24)32(36)40-33)38-31(35)23-9-3-5-11-29(23)41-42-30-12-6-7-17-34-30/h2-19H,1H3
Chemical Name	(6'-methoxy-3-oxospiro[2-benzofuran-1,9'-xanthene]-3'-yl) 2-(pyridin-2-yldisulfanyl)benzoate
HS Tariff Code	2934.99.9001
Storage	Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: This product requires protection from light (avoid light exposure) during transportation and storage.
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

ln Vitro

WSP-1 exhibits fluorescence, with emission wavelength (em) of 515 nm and excitation wavelength (ex) of 465 nm[1]. Advice (Our suggested scenario is shown here. This hypothetical situation should be tailored to your unique requirements; it merely offers suggestions. Fluorescence microscopy for intracellular H2S detection [1]: 1. Grow HASMCs until aggregation reaches roughly 90%. Cells were cultivated on an 8-well culture slide for 24 hours at a density of 30*103 per well, 24 hours prior to the experiment. Time 2. Incubate for 30 minutes at 37°C after replacing the culture medium and adding 400 μL of WSP-1 working solution. 3. Add 20 μL of the H2S donor element at the necessary concentration and swap out the WSP-1 working solution for 380 μL of standard buffer. 4. WSP-1 releases a fluorophore at Ex/Em=465/515 nm in response to H2S. After an hour of incubation and washing with 400 μL DPBS, take the buffer standard. 5. After removing the DPBS, put 200 μL of Bouin solution into each well to fix the cells for ten minutes. Keep Bouin's solution aside and use 400 μL of DPBS to wash the cells twice. 6. To obtain signals, use fluorescence microscopy and fluorescence.

References

[1]. Vascular Effects of H2S-Donors: Fluorimetric Detection of H2S Generation and Ion Channel Activation in Human Aortic Smooth Muscle Cells. Methods Mol Biol. 2019;2007:79-87.

Solubility Data

Solubility (In Vitro)

DMSO : ~25 mg/mL (~42.25 mM)

Solubility (In Vivo)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] *Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.6902 mL	8.4509 mL	16.9019 mL
	5 mM	0.3380 mL	1.6902 mL	3.3804 mL
	10 mM	0.1690 mL	0.8451 mL	1.6902 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.